tag:blogger.com,1999:blog-61810737195156327152024-02-27T06:57:52.548-05:00Democratizing Cell TechnologiesStrategies, tools and technologies for simplifying the incorporation of living cells into tomorrow's products.Jon Rowleyhttp://www.blogger.com/profile/10634518820328346758noreply@blogger.comBlogger52125tag:blogger.com,1999:blog-6181073719515632715.post-21661268977617539852020-01-28T14:26:00.001-05:002020-01-28T14:26:33.033-05:00Our blog has moved!We've integrated our blog into our new website at www.roosterbio.com. Please check it out!Sandy Applebeehttp://www.blogger.com/profile/01351576729478919332noreply@blogger.com0tag:blogger.com,1999:blog-6181073719515632715.post-91295242084153570652020-01-02T15:13:00.001-05:002020-01-02T15:41:35.358-05:00hUC-MSC Exhibit Robust Proliferation in 3D Bioreactor Systems<div class="MsoNormal" style="font-family: "Times New Roman", serif; margin: 0in 0in 0.0001pt; text-align: left;">
<span style="font-family: "calibri" , sans-serif; font-size: 10pt;"><i>Authored by: Joseph Takacs, MS, Research Associate, RoosterBio and Katrina Adlerz, PhD, Scientist, RoosterBio</i><o:p></o:p></span></div>
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<b><span style="font-family: inherit;">Scalable Manufacturing Solution Needed<o:p></o:p></span></b></div>
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<span style="font-family: inherit;"><a href="https://www.blogger.com/null" name="_Hlk23341003"></a>Historically, pre-clinical studies and clinical trials have predominantly used hMSCs derived from bone marrow and adipose tissue sources. However, over the last 10 to 15 years, the number of publications and clinical trials in the regenerative medicine field using hMSCs derived from human umbilical cord (hUC-MSCs) as a raw material has significantly increased (<a href="https://www.ncbi.nlm.nih.gov/pubmed/28488282" style="color: #954f72;">1</a>,<a href="https://www.ncbi.nlm.nih.gov/pubmed/29415771" style="color: #954f72;">2</a>,<a href="http://roosterbio.blogspot.com/2019/09/meeting-growing-needs-of-perinatal.html" style="color: #954f72;">3</a>).<o:p></o:p></span></div>
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<span style="font-family: inherit;"><span style="font-family: inherit;">The commercial hUC-MSC therapies that will follow will require hundreds of millions to billions of cells per lot for effective patient dosing (<a href="http://roosterbio.blogspot.com/2018/12/building-effective-multi-year-process.html" style="color: #954f72;"><span style="background-color: white;">4</span></a>,<a href="https://www.ncbi.nlm.nih.gov/pubmed/29977893" style="color: #954f72;">5</a>). Traditional 2-dimensional (2D) flask expansion platforms are not cost-effective for such large-scale expansion of cell-based therapeutics. However, 3-dimensional (3D), microcarrier-based, bioreactor systems offer a scalable manufacturing platform that can achieve the lot sizes needed. Here, we demonstrate that hUC-MSCs achieve high cell densities in a 3D bioreactor culture system and maintain their critical quality attributes (CQAs) after harvest.</span><o:p></o:p></span></div>
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<b><span style="font-family: inherit;">hUC-MSC Expansion in Scalable Bioreactor Technologies<o:p></o:p></span></b></div>
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<span style="font-family: inherit;"><span style="font-family: inherit;">hUC-MSCs derived from three different donors were cultured in a xeno-free (XF), fed-batch suspension 100mL bioreactor system according to RoosterBio’s bioreactor process recommendations. hUC-MSCs from all three donors reached high cell densities of between 0.8 x 10<sup>6</sup>and 1.2 x 10<sup>6 </sup>cells/mL after five days of culture (Figure 1). Previous experience with bone marrow derived hMSCs suggests that bioreactor systems can be scaled to larger volumes such as 3, 15, and 50L systems with similar or even improved cell growth (<a href="https://www.sciencedirect.com/science/article/abs/pii/S1465324919306152" style="color: #954f72;"><span style="color: windowtext;">6</span></a>).</span><o:p></o:p></span></div>
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<b><span style="font-family: inherit;">Figure 1: hUC-MSCs reach new expansion potential.<o:p></o:p></span></b></div>
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<span style="font-family: inherit;">(A) hUC-MSCs were cultured for five days in a fed-batch bioreactor system with a RoosterReplenish-MSC-XF feed on Day 3. hUC-MSCs reached cell densities of at least 0.8 x 10<sup>6</sup>cells/mL by Day 5. (B) By Day 5, cell-microcarrier aggregation and sampled cell counts indicated that cultures were ready for harvest.<o:p></o:p></span></div>
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<b><span style="font-family: inherit;">hUC-MSCs Robust Functional Capability<o:p></o:p></span></b></div>
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<span style="font-family: inherit;">To ensure these cells maintained hMSC CQAs, hUC-MSCs expanded in 3D bioreactors were compared to hUC-MSCs expanded in 2D flasks in a panel of assays (<a href="http://www.ncbi.nlm.nih.gov/pubmed/16923606" style="color: #954f72;"><span style="background-color: white;">7</span></a>). After expansion in control flasks (2D) or bioreactors (3D), harvested cells were analyzed for: expansion over a subsequent passage, surface marker expression by flow cytometry, immunomodulatory properties, angiogenic cytokine secretion, and trilineage differentiation (Figure 2). While there was donor-to donor variability, hUC-MSCs that were expanded in 3D bioreactors performed comparably to hUC-MSCs that were expanded in 2D flasks. <o:p></o:p></span></div>
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<span style="font-family: inherit;"><span style="font-family: inherit;">This study demonstrates that hUC-MSCs can be expanded in a 3D bioreactor system, reach high cell densities, and maintain their CQAs after harvest. Therefore, hUC-MSCs paired with a bioreactor platform is a system that can be scaled to yield the cell numbers required for product development and commercial therapeutics.</span><span style="color: red;"><o:p></o:p></span></span></div>
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<b><span style="font-family: inherit;">Figure 2: hUC-MSCs maintain critical quality attributes after 3D culture.<o:p></o:p></span></b></div>
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<span style="font-family: inherit;"><span style="font-family: inherit;">Freshly harvested cells were plated into functional assays to determine: (A) expansion capacity over a subsequent passage (B) surface marker expression by flow cytometry (representative donor shown) (C) immunomodulatory properties through the secretion of indoleamine-2,3-dioxygenase (D) angiogenic cytokine secretion and (E) trilineage differentiation through media induction (representative donor shown).</span><b><o:p></o:p></b></span></div>
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<b><span style="font-family: inherit;">References<o:p></o:p></span></b></div>
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<span style="background-color: white;"><span style="font-family: inherit;">1. Davies JE, Walker JT, Keating A. (2017) Concise Review: Wharton's Jelly: The Rich, but Enigmatic, Source of Mesenchymal Stromal Cells. <i>Stem Cells Transl Med.,</i>6(7):1620-1630. <u>https://www.ncbi.nlm.nih.gov/pubmed/28488282</u><o:p></o:p></span></span></div>
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<span style="font-family: inherit;"><span style="background-color: white;">2.</span>Zhao J, Yu G, Cai M, Lei X, Yang Y, Wang Q, Zhai X (2018) Bibliometric analysis of global scientific activity on umbilical cord mesenchymal stem cells: a swiftly expanding and shifting focus<i>. Stem Cell Research & Therapy,</i>9(32). <u>https://www.ncbi.nlm.nih.gov/pubmed/29415771<o:p></o:p></u></span></div>
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<span style="font-family: inherit;">3.<span style="background-color: white; background-position: initial initial; background-repeat: initial initial;">Farrance I (2019) Meeting the growing needs of the perinatal RegenMed Industry: the only Umbilical Cord hMSC (hUC-MSC) system designed for today’s translationally focused research and product development. RoosterBio Blog, 18 September 2019. <u>h</u></span><u>ttp://roosterbio.blogspot.com/2019/09/meeting-growing-needs-of-perinatal.html</u><span style="background-color: white; background-position: initial initial; background-repeat: initial initial;"><o:p></o:p></span></span></div>
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<span style="font-family: inherit;"><span style="background-color: white;">4. Lembong J, Rowley J (2018) Building Effective Multi-Year Process Development Programs: Evolution of Technology Platform Decisions Based on Lot Size. RoosterBio Blog, 15 December 2018. </span><a href="http://roosterbio.blogspot.com/2018/12/building-effective-multi-year-process.html" style="color: #954f72;"><span style="background-color: white; color: black;">http://roosterbio.blogspot.com/2018/12/building-effective-multi-year-process.html</span></a><u><o:p></o:p></u></span></div>
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<span style="font-family: inherit;">5. Olsen TR, Ng KS, Lock LT, Ahsan T, Rowley JA (2018) Peak MSC – are we there yet? <i>Front. Med.,</i>21 June 2018. <a href="https://www.blogger.com/null" name="_Hlk24707087"></a><a href="https://www.ncbi.nlm.nih.gov/pubmed/29977893" style="color: #954f72;"><span style="color: black;">https://www.ncbi.nlm.nih.gov/pubmed/29977893</span></a><o:p></o:p></span></div>
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<span style="font-family: inherit;">6. Kirian RD, Wang D, Takacs J, Tsai A, Cruz K, Rosello F, Cox K, Hashimura Y, Lembong J, Rowley JA, Jung S (2019) Scaling A Xeno-Free Fed-Batch Microcarrier Suspension Bioreactor System From Development to Production Scale for Manufacturing XF hMSCs. <i>Cytotherapy</i>, 2019 May 1;21(5):S71-2.<o:p></o:p></span></div>
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<span style="font-family: inherit;">7. <span style="background-color: white; background-position: initial initial; background-repeat: initial initial;">Dominici M, Le Blanc K, Mueller I, Slaper-Cortenbach I, Marini F, Krause D, Deans R, Keating A, Prockop D, & Horwitz E (2006) Minimal criteria for defining multipotent mesenchymal stromal cells. The International Society for Cellular Therapy position statement. <i>Cytotherapy,</i> 8(4):315-317. </span><a href="http://www.ncbi.nlm.nih.gov/pubmed/16923606" style="color: #954f72;"><span style="background-color: white; color: black;">http://www.ncbi.nlm.nih.gov/pubmed/16923606</span></a></span><span style="background-color: white;"><span style="font-family: inherit;">.</span><span style="font-family: "calibri" , sans-serif; font-size: 11pt;"><o:p></o:p></span></span></div>
Sandy Applebeehttp://www.blogger.com/profile/01351576729478919332noreply@blogger.com0tag:blogger.com,1999:blog-6181073719515632715.post-78276393706266874792019-12-17T16:19:00.001-05:002020-01-08T17:52:23.823-05:00Top 10 Questions Received by RoosterBio Customer Support<table cellpadding="0" cellspacing="0" class="tr-caption-container" style="float: right; text-align: right;"><tbody>
<tr><td style="text-align: center;"><a href="https://blogger.googleusercontent.com/img/b/R29vZ2xl/AVvXsEhRq_JF917lEB0L7IpyKDVqpL3ITUqiRL2777QnOFTC96IPstdyBrhDTTUmKWm4j7qwCEXQqSU1lbGpUL_COSjEmSg_i9uHngJuK1hoyoBzFb-2T5CuZcDBd1snVmEJtPxi2NLXobYYEGjL/s1600/CustomerSupport.jpg" imageanchor="1" style="margin-left: auto; margin-right: auto;"><img border="0" data-original-height="1195" data-original-width="1600" height="149" src="https://blogger.googleusercontent.com/img/b/R29vZ2xl/AVvXsEhRq_JF917lEB0L7IpyKDVqpL3ITUqiRL2777QnOFTC96IPstdyBrhDTTUmKWm4j7qwCEXQqSU1lbGpUL_COSjEmSg_i9uHngJuK1hoyoBzFb-2T5CuZcDBd1snVmEJtPxi2NLXobYYEGjL/s200/CustomerSupport.jpg" width="200" /></a></td></tr>
<tr><td class="tr-caption" style="text-align: center;"><i>Tom Rogers, Director, Global Customer Support <br />and Amy Permenter, Technical Support Specialist</i></td></tr>
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<i>Answered by: Amy Permenter, Technical Support Specialist and Tom Rogers, Director, Global Customer Support</i><br />
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<b>1. Do you have a sales representative in my state or country?</b><br />
Yes! RoosterBio has four outstanding Regional Account Managers:<br />
<ul>
<li><a href="mailto:tim@roosterbio.com">Tim Olsen</a> - Northeast US, Eastern Canada, Europe and Israel</li>
<li><a href="mailto:kmillar@roosterbio.com">Katie Millar</a> - Southeast US, MI, IN and OH</li>
<li><a href="mailto:jdiesselhorst@roosterbio.com">Josh Diesselhorst </a>- Midwest US, including TX, and Central Canada</li>
<li><a href="mailto:mlim@roosterbio.com">Maya Lim</a> - Western US, Western Canada, Asia and Australia</li>
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<a href="https://blogger.googleusercontent.com/img/b/R29vZ2xl/AVvXsEjgtFPJQRpqTFuaa8hi6k5_vw5c89xHhIibtXy5ceiopbJolJttUQHlTdpK472lmInwQ0-Qeleo__W7IJefe_qhwp570qFqCueRSN-c_TXNUPws9yAOaNFf_2SQekCDwo-TdslYLYJ3AhCc/s1600/Screen+Shot+2019-12-12+at+2.02.23+PM.png" imageanchor="1" style="margin-left: 1em; margin-right: 1em;"><img border="0" data-original-height="935" data-original-width="1600" height="371" src="https://blogger.googleusercontent.com/img/b/R29vZ2xl/AVvXsEjgtFPJQRpqTFuaa8hi6k5_vw5c89xHhIibtXy5ceiopbJolJttUQHlTdpK472lmInwQ0-Qeleo__W7IJefe_qhwp570qFqCueRSN-c_TXNUPws9yAOaNFf_2SQekCDwo-TdslYLYJ3AhCc/s640/Screen+Shot+2019-12-12+at+2.02.23+PM.png" width="640" /></a></div>
RoosterBio has two talented Field Application Scientists (FAS), <a href="mailto:xxu@roosterbio.com">Xuan Xu </a>and <a href="mailto:joseph@roosterbio.com">Joe Takacs</a>, who can assist in the field with technical questions. <a href="mailto:jgetz@roosterbio.com">John Getz </a>is RoosterBio's Government Account Manager and can assist with government-related questions. If you are unsure of who to contact, please contact <a href="mailto:tom@roosterbio.com">Tom Rogers</a>.<br />
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<b>2. How do I request a quote for your products?</b></div>
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RoosterBio can provide a quote for any purchase. Contact your local Regional Account Manager or <a href="mailto:info@roosterbio.com">Customer Support</a> at 240-831-4914.</div>
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<b>3. I'm ready to make a purchase! What's next?</b></div>
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Great! We have three different ways you can purchase our products:</div>
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<ul>
<li>Website - <a href="http://www.roosterbio.com/">www.roosterbio.com</a></li>
<li>Purchase Order - submit to your Regional Account Manager or <a href="mailto:info@roosterbio.com">Customer Support</a></li>
<li>Credit Card Purchase - submit a Credit Card Authorization Form (obtained from <a href="mailto:info@roosterbio.com">Customer Support</a>) to your Regional Account Manager or Customer Support</li>
</ul>
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<b>4. How do you ship your products and how long does it take to arrive?</b></div>
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RoosterBio ships all over the world - 26 countries ... and counting!</div>
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<ul>
<li>For local orders in <u>Maryland</u>, we use a courier service for same day delivery. </li>
<li>For <u>US</u> orders, we use FedEx overnight priority delivery. This is next day delivery and should arrive by 12:00pm.</li>
<li>For orders to <u>Canada and Europe,</u> we use FedEx and it usually takes 2-4 days depending on customs clearance.</li>
<li>For orders to <u>Asia and Australia</u>, we use FedEx and it usually takes 3-5 days depending on customs clearance. Note: we have used Expeditors and World Courier for some shipments to Asia.</li>
</ul>
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<b>5. What documentation is sent with the order?</b></div>
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<ul>
<li>For <b>cell orders</b>, you will receive an Expansion Protocol, QC Brief, Certificate of Analysis and Cell Product Insert.</li>
<li>For <b>media orders</b>, you will receive a Certificate of Analysis and the Media Product Insert.</li>
<li>For <b>international customers, </b>you will also receive a Material Safety Data Sheet for all products ordered. Depending on which country you are in, you could also receive a Certificate of Origin.</li>
</ul>
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<b>6. The products arrived! Now what do I do?</b></div>
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<ul>
<li>Place the cells in liquid nitrogen storage</li>
<li>For the RoosterNourish™ Media:</li>
<ul>
<li>Store the RoosterBasal™ in 2-8°C refrigerator (protect it from light)</li>
<li>Store the RoosterBooster™ in -20°C freezer</li>
</ul>
<li>Store the RoosterReplenish™ in -20°C freezer</li>
<li>Store the EV Boost™ in a 2-8°C refrigerator (protect it from light)</li>
<li>Store the RoosterCollect™ in a 2-8°C refrigerator (protect it from light)</li>
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<b>7. I ordered my cells and media, but when my order arrived, I couldn't find the cells. Help!</b></div>
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Do not worry! RoosterBio sometimes uses a nest configuration in one large box when shipping cells and media together. Inside the large box is the RoosterBasal™ on cold bricks, along with a smaller box. The smaller box is layered with dry ice, the RoosterBooster™ and the cell vial. Please make sure to remove the RoosterBooster™ and dig deeper into the box for the cell vial.</div>
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<b>8. I'm ready to use the RoosterNourish™ Media Kit. What do I do? How long can I use the media?</b></div>
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Bring the Media Kit to room temperature. In a hood, pipet the 10mL RoosterBooster™ into the 500mL RoosterBasal™. Once combined, store the media at 2-8°C for 2 weeks, ensuring that it is protected from light.</div>
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<b>9. Do I need to heat inactivate the media?</b></div>
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No, you do not.</div>
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<b>10. Can I request a specific lot of cells?</b></div>
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Absolutely! RoosterBio has a donor grid that includes age, gender, population doubling level (PDL), differentiation and flow markers. Contact <a href="mailto:info@roosterbio.com">Customer Support</a> for the donor grid.<br />
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Sandy Applebeehttp://www.blogger.com/profile/01351576729478919332noreply@blogger.com0tag:blogger.com,1999:blog-6181073719515632715.post-65648086419414489842019-11-26T15:11:00.003-05:002020-01-02T15:33:57.472-05:00Generating MSC-EVs With A Scalable Manufacturing System<div class="MsoNormal" style="color: black; letter-spacing: normal; margin: 0in 0in 0.0001pt; text-align: justify; text-decoration: none; text-indent: 0px; text-transform: none; white-space: normal; word-spacing: 0px;">
<span style="font-family: inherit; font-size: 11pt;"><i>Authored by: Katrina Adlerz, PhD, Scientist, Analytical, Process & Product Development, RoosterBio</i></span></div>
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<b><span style="font-family: inherit; font-size: 11pt;">Scalability of EV Manufacturing is a Major Challenge<o:p></o:p></span></b></div>
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<span style="font-family: inherit;"><span style="font-family: "calibri" , sans-serif; font-size: 11pt;">Previously, we discussed the <a href="http://roosterbio.blogspot.com/2019/11/msc-evs-emerge-as-clinical-therapies.html">emergence of MSC extracellular vesicles (EVs) as clinical therapies</a>. However, a <b>critical barrier </b>in the development of MSC-EVs as a commercial therapy is <b>generating the large amount of EVs </b><b>that will be required per dose </b></span><span style="font-family: "calibri" , sans-serif; font-size: 11pt;">(1)</span><span style="font-family: "calibri" , sans-serif; font-size: 11pt;">. A recent review estimated that the number of exosomes released from 2 million MSCs in 48 hours is equivalent to a single dose for a rodent </span><span style="font-family: "calibri" , sans-serif; font-size: 11pt;">(2)</span><span style="font-family: "calibri" , sans-serif; font-size: 11pt;">, suggesting that similar to cell therapy, <a href="http://roosterbio.blogspot.com/2018/12/building-effective-multi-year-process.html">billions of cells will be required to manufacture an EV commercial therapy</a>. In a recent survey, however, the majority of those working with EVs were working with less than 100mL of sample </span><span style="font-family: "calibri" , sans-serif; font-size: 11pt;">(3) </span><span style="font-family: "calibri" , sans-serif; font-size: 11pt;">indicating the lack of scalable manufacturing processes in the field. We have identified three keys that we believe are necessary to enable successful manufacturing of EVs for clinical therapies:</span></span></div>
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<li><span style="font-family: inherit; text-indent: -0.25in;">Generating the billions of cells needed with a scalable manufacturing process</span></li>
<li><span style="font-family: inherit; text-indent: -0.25in;">Increasing the number of EVs generated per cell to maximize productivity</span></li>
<li><span style="font-family: inherit; text-indent: -0.25in;">Optimizing downstream EV purification to increase concentration with minimal processing loss (to be addressed in future blog)</span></li>
<li><span style="font-family: inherit; text-indent: -0.25in;">GMP quality supply chain takes years to develop. Starting with the right materials is critical.</span></li>
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<a href="https://blogger.googleusercontent.com/img/b/R29vZ2xl/AVvXsEiDh9ueWA1wPegGzCHomUUcRzD_znjQV36I560IoPzy2oLjSvC7KeFIcclQscx5u63KPceZYqrJ1_PduFUbvU_5Vn9wk2AIc7G3ZyigWCbyMingOHlcBtR8HYBAFQrzbQkNWLl4fP6dHm24/s1600/Screen+Shot+2019-11-26+at+2.59.17+PM.png" imageanchor="1" style="clear: right; float: right; margin-bottom: 1em; margin-left: 1em;"><span style="font-family: inherit;"><img border="0" data-original-height="476" data-original-width="616" height="246" src="https://blogger.googleusercontent.com/img/b/R29vZ2xl/AVvXsEiDh9ueWA1wPegGzCHomUUcRzD_znjQV36I560IoPzy2oLjSvC7KeFIcclQscx5u63KPceZYqrJ1_PduFUbvU_5Vn9wk2AIc7G3ZyigWCbyMingOHlcBtR8HYBAFQrzbQkNWLl4fP6dHm24/s320/Screen+Shot+2019-11-26+at+2.59.17+PM.png" width="320" /></span></a></div>
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<b><span style="font-family: inherit; font-size: 11pt;">More Cells Enable Production of More EVs<o:p></o:p></span></b></div>
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<span style="font-family: inherit; font-size: 11pt;">The first critical challenge to generating the EVs needed for product development and clinical therapies is <b>growing the necessary numbers of cells</b>. RoosterBio (RBI) high-volume cell formats and paired bioprocess growth medium are engineered for scalable manufacturingto address this bottleneck. Figure 1 illustrates the ability of RBI systems to generate millions to billions of MSCs, which produce trillions of EVs, in an 8 to 10 day process. Our starting Working Cell Bank vial format and culture paradigm decrease manufacturing time and are scalable to yield the number of EVs required for clinical translation.<o:p></o:p></span></div>
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<b><span style="font-family: inherit; font-size: 11pt;">Scalable Process for MSC-EV Manufacturing<o:p></o:p></span></b></div>
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<span style="font-family: inherit; font-size: 11pt;">We recently introduced RoosterCollect<sup>TM</sup>-EV, a low-particle medium that is engineered for EV collection. This medium is designed to be complementary to RoosterBio MSCs and the RoosterBio bioprocess growth medium RoosterNourish<sup>TM</sup>. Together, these RBI products create a <b>complete system for efficient cell growth and EV collection</b>. </span></div>
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<a href="https://blogger.googleusercontent.com/img/b/R29vZ2xl/AVvXsEh20Kdei0AKaGj0TU6aKJQ5tw5uU56UOYKX9GB6eD38A6w9Z-xuhXabLwbVz6MIVn5WzoEdeXPamDmpOQlPdvjwqJnW342MQGfThxozEtvgvFm2cqmsnISSpYTZyROD3tFwbh_omNYRjMMl/s1600/Screen+Shot+2019-11-26+at+3.00.19+PM.png" imageanchor="1" style="clear: right; float: right; margin-bottom: 1em; margin-left: 1em;"><span style="font-family: inherit;"><img border="0" data-original-height="430" data-original-width="632" height="217" src="https://blogger.googleusercontent.com/img/b/R29vZ2xl/AVvXsEh20Kdei0AKaGj0TU6aKJQ5tw5uU56UOYKX9GB6eD38A6w9Z-xuhXabLwbVz6MIVn5WzoEdeXPamDmpOQlPdvjwqJnW342MQGfThxozEtvgvFm2cqmsnISSpYTZyROD3tFwbh_omNYRjMMl/s320/Screen+Shot+2019-11-26+at+3.00.19+PM.png" width="320" /></span></a></div>
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<span style="font-family: inherit;"><span style="font-family: inherit; font-size: 11pt;">Using these products, the optimized process (</span><span style="font-family: "calibri" , sans-serif; font-size: 11pt;"><span style="font-family: inherit;">shown in Figure 2) is: </span><o:p></o:p></span></span></div>
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<span style="font-family: inherit; font-size: 11pt;">1) Expand MSCs in RoosterNourish until at least 80% confluency <o:p></o:p></span></div>
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<span style="font-family: inherit; font-size: 11pt;">2) Switch to RoosterCollect-EV <o:p></o:p></span></div>
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<span style="font-family: inherit; font-size: 11pt;">3) Collect the conditioned medium<o:p></o:p></span></div>
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<span style="font-family: inherit; font-size: 11pt;">RoosterCollect-EV supports EV collection for at least two days with increasing particle concentration in the conditioned medium and collected particles in the size range expected for EVs (Figure 3).</span><br />
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<a href="https://blogger.googleusercontent.com/img/b/R29vZ2xl/AVvXsEibdwINKyNrbzsQF0d8g0mRDiorDWc7oxHqgA9PndfofcJBEL3GbQtRMp2UztnFMEz_nTC_HdD2zhBQxqOl5t9RO9EU9BbJ0K3OOs1EANZ1g1AEy5gvlL012vJjD7V4Pw8pa2ylPE2VqUts/s1600/Screen+Shot+2019-11-26+at+3.04.15+PM.png" imageanchor="1" style="margin-left: 1em; margin-right: 1em;"><span style="font-family: inherit;"><img border="0" data-original-height="462" data-original-width="1194" height="244" src="https://blogger.googleusercontent.com/img/b/R29vZ2xl/AVvXsEibdwINKyNrbzsQF0d8g0mRDiorDWc7oxHqgA9PndfofcJBEL3GbQtRMp2UztnFMEz_nTC_HdD2zhBQxqOl5t9RO9EU9BbJ0K3OOs1EANZ1g1AEy5gvlL012vJjD7V4Pw8pa2ylPE2VqUts/s640/Screen+Shot+2019-11-26+at+3.04.15+PM.png" width="640" /></span></a></div>
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<w:wrap type="square"><w:wrap type="square"><w:wrap type="topAndBottom"><w:wrap type="square"><span style="font-family: inherit; font-size: 11pt;">This process works for both 2D flask culture and 3D bioreactor culture. Bioreactor culture allows for even greater MSC and EV yields with reductions in cost, labor, and time <a href="https://info.roosterbio.com/roosterbio-isct2019-follow-up-0-0-2">(see our poster presented at ISCT 2019)</a>. Also, conditioned media in 3L and 15L bioreactor culture had greater particle concentration compared to the 2D flask system (Figure 4), which is at least partly explained by the increased cell density we can achieve in bioreactors <a href="https://info.roosterbio.com/roosterbio-isev2019-follow-up-sm">(see our poster presented at ISEV 2019)</a>. <o:p></o:p></span></w:wrap></w:wrap></w:wrap></w:wrap><br />
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<b><span style="font-family: inherit; font-size: 11pt;">Increased Productivity for More EVs<o:p></o:p></span></b></div>
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<w:wrap type="square"><w:wrap type="square"><w:wrap type="topAndBottom"><w:wrap type="square"><w:wrap type="square"><span style="font-family: inherit; font-size: 11pt;">A complementary strategy to a scalable cell manufacturing process is <b>increasing the EV productivity </b>(i.e. the number of EVs produced per cell). Recently-introduced EV Boost<sup>TM </sup>is a medium supplement that is designed as a tunable addition to RoosterCollect-EV medium. Depending on the number of EVs required, EV Boost can increase particle yield up to 5x and dramatically shorten collection times (Figure 5).<o:p></o:p></span></w:wrap></w:wrap></w:wrap></w:wrap></w:wrap><br />
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<b><span style="font-family: inherit; font-size: 11pt;">What’s Next in Scalable EV Production?<o:p></o:p></span></b></div>
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<span style="font-family: inherit; font-size: 11pt;">Optimizing EV yield will become increasingly important as EVs move to clinical therapies and EVs from billions of cells are required to satisfy dose requirements. RoosterBio’s complete system for MSC-EVs generates millions to billions of cells in 2D or bioreactor culture and trillions of EVs, this system also provides a scalable platform for increasing EV production. This, combined with optimized scalable downstream purification (a third key challenge in EV manufacturing, and subject of a future blog), will enable the success of EVs as clinical therapies. </span><br />
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<span style="font-family: "calibri" , sans-serif; font-size: 11pt;">Rapid translation/transition of to cGMP production will drive the future of scalable EV production for years to come.<o:p></o:p></span></span></div>
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<b><span style="font-family: inherit; font-size: 11pt;">References:<o:p></o:p></span></b></div>
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<span style="font-family: inherit; font-size: 11pt;">1. Colao IL, Corteling R, Bracewell D, Wall I. Manufacturing Exosomes: A Promising Therapeutic Platform. Trends Mol Med. 2018;24(3):242-56.<o:p></o:p></span></div>
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<span style="font-family: inherit; font-size: 11pt;">2. Phinney DG, Pittenger MF. Concise Review: MSC-Derived Exosomes for Cell-Free Therapy. Stem Cells. 2017;35(4):851-8.</span></div>
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<span style="font-family: "calibri" , sans-serif; font-size: 11pt;"><span style="font-family: inherit;">3. Gardiner C, Di Vizio D, Sahoo S, Théry C, Witwer KW, Wauben M, et al. Techniques used for the isolation and characterization of extracellular vesicles: results of a worldwide survey. J Extracell Vesicles. 2016;5:32945.</span><span style="font-family: calibri, sans-serif; font-size: 11pt;"><o:p></o:p></span></span></div>
Sandy Applebeehttp://www.blogger.com/profile/01351576729478919332noreply@blogger.com0tag:blogger.com,1999:blog-6181073719515632715.post-7596823102769418952019-11-01T15:38:00.000-04:002020-01-02T15:34:17.730-05:00MSC-EVs Emerge as Clinical Therapies<div class="MsoNormal" style="font-family: "Times New Roman", serif; margin: 0in 0in 0.0001pt; text-align: left;">
<span style="font-family: "calibri" , sans-serif; font-size: 10pt;"><i>Authored by: Katrina Adlerz, PhD, Scientist, RoosterBio and Divya Patel, PhD, Scientist, RoosterBio<o:p></o:p></i></span></div>
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<b><span style="font-family: "calibri" , sans-serif; font-size: 11pt;">MSC-EVs Emerge as a Cell-Free Therapy<o:p></o:p></span></b><br />
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<span style="font-family: "calibri" , sans-serif; font-size: 11pt;">Extracellular vesicle (EV) interest continues to increase as more evidence emerges about the ability of these lipid-bilayer membrane vesicles to elicit specific responses from recipient cells. EVs are secreted by most known cell types, including MSCs. Recently, many effects of MSC-based therapeutics have been attributed to their paracrine factors which includes MSC-derived EVs </span><span style="font-family: "calibri" , sans-serif; font-size: 11pt;">(1, 2)</span><span style="font-family: "calibri" , sans-serif; font-size: 11pt;">. In particular, MSC-derived EVs have been shown to recapitulate <b>therapeutic effects of MSCs </b>in graft-versus-host disease </span><span style="font-family: "calibri" , sans-serif; font-size: 11pt;">(3)</span><span style="font-family: "calibri" , sans-serif; font-size: 11pt;">and myocardial ischemia </span><span style="font-family: "calibri" , sans-serif; font-size: 11pt;">(4)</span><span style="font-family: "calibri" , sans-serif; font-size: 11pt;">, among others. Moreover, EVs derived from MSCs benefit from MSCs’ well-defined safety profile, with MSCs having been used in over 900 clinical trials. Given their therapeutic potential, EVs are on the rise as a novel clinical therapy for a broad range of applications. This interest is reflected in the high number of peer-reviewed publications in the past 10 years mentioning EVs (over 15,000), with 700 specifically on MSC-EVs (PubMed Search Results Oct 2019) and the <a href="http://roosterbio.blogspot.com/2019/07/advances-in-clinical-translation-and.html">larger presence of EVs at cell therapy conferences</a>.<o:p></o:p></span></div>
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<b><span style="font-family: "calibri" , sans-serif; font-size: 11pt;">MSC-EVs as Drug Delivery Vehicles<o:p></o:p></span></b><br />
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<span style="font-family: "calibri" , sans-serif; font-size: 11pt;">In addition to their use as a cell-free therapy, there is also significant interest in using EVs as <b>drug delivery vehicles</b>. EVs are natural carriers of bioactive cargo such as proteins and RNA, which are protected by the lipid-bilayer membrane. Research efforts have focused on both exogenous loading of biological cargo and manipulating parent cells to engineer vesicles that contain cargo of interest. <o:p></o:p></span></div>
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<b><span style="font-family: "calibri" , sans-serif; font-size: 11pt;">RoosterBio EVs</span></b><br />
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<w:wrap style="font-family: -webkit-standard;" type="topAndBottom"><span style="font-family: "calibri" , sans-serif; font-size: 11pt;">RoosterBio MSC-EVs were evaluated based on a guidance for defining EVs, as published by the <a href="https://www.isev.org/">International Society for Extracellular Vesicles</a> </span><span style="font-family: "calibri" , sans-serif; font-size: 11pt;">(5)</span><span style="font-family: "calibri" , sans-serif; font-size: 11pt;">. Particles collected from the conditioned medium of RoosterBio MSCs are in the EV size range, contain expected proteins and small RNAs, and have bioactivity in a wound healing assay (Figure 1).<o:p></o:p></span></w:wrap></div>
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<w:wrap style="font-family: -webkit-standard;" type="topAndBottom"><b style="font-family: "Times New Roman", serif; font-size: 13.333333015441895px; font-style: italic;"><span style="font-family: "calibri" , sans-serif; font-style: normal;">Figure </span></b><b style="font-family: "Times New Roman", serif; font-size: 13.333333015441895px; font-style: italic;"><span style="font-family: "calibri" , sans-serif; font-style: normal;">1</span></b><b style="font-family: "Times New Roman", serif; font-size: 13.333333015441895px; font-style: italic;"><span style="font-family: "calibri" , sans-serif; font-style: normal;"> A </span></b><span style="font-family: "calibri" , sans-serif; font-size: 13.333333015441895px;">Particles collected from RoosterBio MSCs have diameters in the size range of 50 to 250 nm as measured by Nanosight and TEM. <b>B </b>Western Blot shows EVs express expected proteins: ALIX, TSG101, CD63, CD9, and CD81. <b>C </b>Collected EVs contain primarily small RNA. <b>D </b>Conditioned medium has bioactivity in a standard in vitro wound healing scratch test.</span></w:wrap><br />
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<b style="text-align: justify;"><span style="font-family: "calibri" , sans-serif; font-size: 11pt;">Challenges Facing the Field</span></b><br />
<span style="font-family: calibri, sans-serif; font-size: 11pt;"><br /></span>
<span style="font-family: calibri, sans-serif; font-size: 11pt;">While EVs hold much promise as a cell-free therapy or drug delivery vehicle, there are some key challenges in the translation of successful EV therapies, including generating the needed number of EVs. In our next blog we will discuss some of these key challenges that need to be addressed to enable the success of EV therapies and RoosterBio’s progress in meeting the needs for EV product development, clinical trials, and commercial therapies. </span><br />
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<b><span style="font-family: "calibri" , sans-serif; font-size: 11pt;"><i>References</i><o:p></o:p></span></b></div>
<div class="EndNoteBibliography" style="font-family: "Times New Roman", serif; margin: 0in 0in 0.0001pt;">
<span style="font-family: "calibri" , sans-serif; font-size: 11pt;">1. Phinney DG, Pittenger MF. Concise Review: MSC-Derived Exosomes for Cell-Free Therapy. Stem Cells. 2017;35(4):851-8.<o:p></o:p></span></div>
<div class="EndNoteBibliography" style="font-family: "Times New Roman", serif; margin: 0in 0in 0.0001pt;">
<span style="font-family: "calibri" , sans-serif; font-size: 11pt;">2. Caplan AI, Correa D. The MSC: an injury drugstore. Cell Stem Cell. 2011;9(1):11-5.<o:p></o:p></span></div>
<div class="EndNoteBibliography" style="font-family: "Times New Roman", serif; margin: 0in 0in 0.0001pt;">
<span style="font-family: "calibri" , sans-serif; font-size: 11pt;">3. Kordelas L, Rebmann V, Ludwig AK, Radtke S, Ruesing J, Doeppner TR, et al. MSC-derived exosomes: a novel tool to treat therapy-refractory graft-versus-host disease. Leukemia. 2014;28(4):970-3.<o:p></o:p></span></div>
<div class="EndNoteBibliography" style="font-family: "Times New Roman", serif; margin: 0in 0in 0.0001pt;">
<span style="font-family: "calibri" , sans-serif; font-size: 11pt;">4. Lai RC, Arslan F, Lee MM, Sze NS, Choo A, Chen TS, et al. Exosome secreted by MSC reduces myocardial ischemia/reperfusion injury. Stem Cell Res. 2010;4(3):214-22.</span></div>
<div class="EndNoteBibliography" style="font-family: "Times New Roman", serif; margin: 0in 0in 8pt;">
<span style="font-family: "calibri" , sans-serif; font-size: 11pt;">5. Lötvall J, Hill AF, Hochberg F, Buzás EI, Di Vizio D, Gardiner C, et al. Minimal experimental requirements for definition of extracellular vesicles and their functions: a position statement from the International Society for Extracellular Vesicles. J Extracell Vesicles. 2014;3:26913.<o:p></o:p></span></div>
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Sandy Applebeehttp://www.blogger.com/profile/01351576729478919332noreply@blogger.com0tag:blogger.com,1999:blog-6181073719515632715.post-78587652553762890522019-09-18T11:14:00.005-04:002019-09-24T11:52:39.094-04:00Meeting the growing needs of the perinatal RegenMed Industry: the only Umbilical Cord hMSC (hUC-MSC) system designed for today’s translationally focused research and product development<i><span style="color: #444444; font-family: "arial" , "helvetica" , sans-serif;">A<span style="color: #444444;">uthored by Iain Farrance, PhD, Technical Marketing Associate</span></span></i><br />
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<b style="mso-bidi-font-weight: normal;"><span style="color: #444444; font-family: "arial" , "helvetica" , sans-serif;"><span style="font-family: "helvetica neue" , "arial" , "helvetica" , sans-serif;">I</span><span style="font-family: "helvetica neue" , "arial" , "helvetica" , sans-serif;"><span style="font-family: "helvetica neue" , "arial" , "helvetica" , sans-serif;">ntroduction</span></span></span></b><br />
<span style="color: #444444;"><span style="font-size: 11pt;"><span style="font-family: "arial" , "helvetica" , sans-serif;"><br /></span></span><span style="font-family: "arial" , "helvetica" , sans-serif;"><span style="font-size: 11pt;">Human mesenchymal stromal cells (hMSC) are considered the <a href="http://roosterbio.blogspot.com/2014/03/mesenchymal-stem-cells-workhorse-of.html">"workhorse" of Regenerative Medicine</a> (RegenMed).</span><span style="font-size: 11pt;"> </span><span style="font-size: 11pt;">hMSC are a critical starting material in a growing variety of established and emerging RegenMed products, including cellular therapies, cell-based gene therapies, hMSC-derived extracellular vesicles (EVs), and bioprinted engineered tissues (Olsen, 2018). Accordingly, there have been greater than 100 clinical trials initiated each year since 2011 using hMSC from various sources (database purchased from celltrials.org) across a host of indications and therapeutic strategies </span><i style="font-size: 11pt;">(<a href="https://clinicaltrials.gov/">clinical trials.gov</a>)</i><span style="font-size: 11pt;">.</span><span style="font-size: 11pt;"> </span><span style="font-size: 11pt;">hMSC have benefitted from having an excellent safety profile, and there have been nine (9) products approved globally over the last 10 years. The growth in use of hMSC in a variety of product types has created the opportunity to standardize the supply chain and provide economies of scale for a rapidly growing industry. RoosterBio was founded to industrialize and standardize the RegenMed supply chain and to radically simplify the incorporation of living cells into therapeutic product development.</span><span style="font-size: 11pt;"> </span><span style="font-size: 11pt;">Our goal is to have the same impact on the RegenMed industry that Intel had on the computer industry.</span></span></span></div>
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<span style="font-family: "arial" , "helvetica" , sans-serif;"><span style="color: #444444; font-family: "arial" , "helvetica" , sans-serif; font-size: 11pt;">Use of </span><span style="font-size: 11pt;"><span style="color: #444444; font-family: "arial" , "helvetica" , sans-serif;">Human Umbilical Cord-derived MSCs (hUC-MSC) in research and clinical
trials (CT) has grown rapidly over the last 10 to 15 years with quickest
adoption in APAC (Figure 1, Davies, 2017; Zhao, 2018; Moll, 2019). hUC-MSC
publications per year increased 19-fold increase from 2006 to 2016 (Zhao, 2018).
CT with hUC-MSC have shown a similar growth pattern as publications. hUC-MSC
are the second most used hMSC type in CT (Moll, 2019) and 178 CT using hUC-MSC
were registered, are ongoing, or were completed between 2007 and 2017 (Couto,
2019). In fact, >30% of hMSC trials registered in 2019 use hUC-MSC as the
cell source. <span style="mso-spacerun: yes;"> </span>These drive the need for
hUC-MSC to use in product development. Until now, IP surrounding hUC-MSC has
been a primary roadblock to the widespread adoption of hUC-MSC. We have collaborated
with leaders in Wharton’s Jelly/umbilical cord hMSC at <a href="http://www.verypowerfulbiology.com/">Tissue RegenerationTherapeutics Inc. </a>(TRT) and have brought to market a complete bioprocess cell
and media system. RoosterBio’s hUC-MSC are available for licensing and are provided
in scalable formulations and cGMP compatible processes that enable anyone to
obtain hUC-MSC in numbers needed for incorporation into RegenMed product
development.</span><span style="color: black; font-family: "arial" , sans-serif;"><o:p></o:p></span></span></span></div>
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<a href="https://blogger.googleusercontent.com/img/b/R29vZ2xl/AVvXsEjDBvT67Wcuk0Q02sFtwbsX4xmEjKkYdotxD4r_aqn3tVHv2qz58ZW8VDqZw2Dd5GXC0TkiS-JaLhm0IQTMysl1c5PSh8qAuf-2_f6jMamoAQgtqMTqZV6ZrwHt1wA5kc7O59uKB4GRZ8kc/s1600/Fig+1.jpeg" imageanchor="1" style="margin-left: 1em; margin-right: 1em; text-align: right;"><span style="font-family: "arial" , "helvetica" , sans-serif;"><img border="0" data-original-height="409" data-original-width="791" height="330" src="https://blogger.googleusercontent.com/img/b/R29vZ2xl/AVvXsEjDBvT67Wcuk0Q02sFtwbsX4xmEjKkYdotxD4r_aqn3tVHv2qz58ZW8VDqZw2Dd5GXC0TkiS-JaLhm0IQTMysl1c5PSh8qAuf-2_f6jMamoAQgtqMTqZV6ZrwHt1wA5kc7O59uKB4GRZ8kc/s640/Fig+1.jpeg" width="640" /></span></a></div>
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<span style="color: #444444; font-family: "arial" , "helvetica" , sans-serif;"><span style="font-family: "arial" , "helvetica" , sans-serif;"><span style="font-size: 11pt;">Until now
RoosterBio has paired our batch (2D) and fed-batch (3D bioreactor) bioprocess
media systems with hMSC from two sources: adipose-derived (hAD-MSC) and bone
marrow-derived (hBM-MSC and xeno-free (XF) hBM-MSC). </span><span style="font-size: 11pt;">RoosterBio’s launch of our XF hUC-MSC (<a href="https://www.roosterbio.com/collections/high-volume-hmscs/products/roostervial-huc-msc-10m">RoosterVial™-hUC-MSC-XF</a>)
introduces the first umbilical cord-derived hMSC in the North American and
worldwide market designed to meet the quality and volume needs of today’s
translationally focused cell therapy product developers. For RoosterBio’s hMSC
product lines see </span></span><a href="https://www.roosterbio.com/collections/high-volume-hmscs" style="font-family: arial, helvetica, sans-serif;"><span style="font-size: 11.0pt;">here</span></a><span style="font-family: "arial" , "helvetica" , sans-serif; font-size: 11pt;">. </span><span style="font-family: "arial" , "helvetica" , sans-serif; font-size: 11pt;"><o:p></o:p></span></span></div>
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<span style="color: #444444; font-family: "arial" , "helvetica" , sans-serif;"><span style="font-size: 11pt;">RoosterBio’s </span><span style="font-size: 11pt;">RoosterVial-hUC-MSC-XF
and <a href="https://www.roosterbio.com/collections/bioprocess-media/products/hmsc-high-performance-media-kit-xf-kt-016">RoosterNourish™-MSC</a> cell and medium</span><span style="font-size: 11pt;"> </span><span style="font-size: 11pt;">bioprocess system</span><span style="font-size: 11pt;"> has several key advantages over the limited
number of suppliers of perinatal hMSC. Being XF, and manufactured with
RoosterBio’s existing cGMP compatible processes, our system </span><span style="font-size: 11pt;">is the only hUC-MSC commercially available with a clear line of sight to
clinical translation. </span><span style="font-size: 11pt;"><span style="mso-spacerun: yes;"> </span></span><span style="font-size: 11pt;">Additionally</span><span style="font-size: 11pt;">, other suppliers (a) provide low cell number
vials at a high price per M cells, (b) supply serum-based cells, or (c) require
specialized, non-scalable culture vessels.<span style="mso-spacerun: yes;">
</span>Finally</span><span style="font-size: 11pt;">, RoosterBio provides first in class
characterization of hMSC key quality attributes (PDL, identity, expansion
potential) and functional assays (cytokine secretion, trilineage
differentiation, immunomodulation).</span></span></div>
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<b style="mso-bidi-font-weight: normal;"><span style="color: #444444; font-family: "arial" , "helvetica" , sans-serif; font-size: 11.0pt;">Results<o:p></o:p></span></b></div>
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<span style="color: #444444; font-family: "arial" , "helvetica" , sans-serif;"><span style="font-size: 11pt;">For a standardized,
high-quality hUC-MSC we put in place stringent design requirements, accounting
for expected donor-to-donor variability, that incorporate hUC-MSCs’ known high
expansion rates and address the ISCT criteria for hMSC </span><span style="font-size: 11pt;">(Carmen, 2012; </span><span style="font-size: 11pt;">Dominici, 2006; Krampera, 2013; Bravery,
2013, </span><span style="font-size: 11pt;">Davies, 2017). <o:p></o:p></span></span></div>
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<span style="color: #444444; font-family: "arial" , "helvetica" , sans-serif;"><span style="font-size: 11.0pt;">As shown below RoosterBio’s
hUC-MSC meet these stringent criteria. Our hUC-MSC, </span><span style="font-size: 11.0pt;">RoosterVial-hUC</span><span style="font-size: 11.0pt;">, (1) have very high expansion rates in
a 2D batch culture, (2) meet ISCT criteria for MSC, (a) cell identity, (b) immunomodulation
and (c) trilineage differentiation, (3) are characterized for functional potency
(angiogenic cytokine secretion), and (4) </span><span style="font-size: 11.0pt;">have unprecedented, quantified productivity
metrics (Carmen, 2012; </span><span style="font-size: 11.0pt;">Dominici,
2006; Krampera, 2013; Bravery, 2013)</span><span style="font-size: 11.0pt;">. <o:p></o:p></span></span></div>
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<span style="color: #444444; font-size: 11.0pt;"><span style="font-family: "arial" , "helvetica" , sans-serif;">Unlike other hMSC
vendors, we perform our quality assays at the PDL (<i style="mso-bidi-font-style: normal;">i.e.</i> level of expansion) that we recommend our customers use our
cells. So, all assays presented here were performed with RoosterBio hUC-MSC
that have been expanded for 2 passages (~10 PDL) from product vials (RoosterVial™-hUC-10M-XF,
C43002 or RoosterVial™-hUC-10M-XF, C43002) in RoosterNourish™-MSC-XF medium,
following RoosterBio’s process recommendations. </span><span style="font-family: "arial" , "helvetica" , sans-serif;"><o:p></o:p></span></span></div>
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<span style="color: #444444; font-family: "arial" , "helvetica" , sans-serif;"><b style="font-family: arial, helvetica, sans-serif;"><i style="mso-bidi-font-style: normal;"><span style="font-size: 11.0pt;">Cell expansion.</span></i></b><i style="font-family: arial, helvetica, sans-serif;"><span style="font-size: 11.0pt;"> </span></i><span style="font-family: "arial" , "helvetica" , sans-serif;"><span style="font-size: 11pt;">A key characteristic
of RoosterBio hMSC bioprocess systems is rapid cell expansion. For example,
RoosterBio’s hBM-MSC-XF in RoosterNourish™-MSC-XF are guaranteed for >10-fold
expansion within 5-7 days. <span style="color: black; mso-themecolor: text1;">See </span></span><a href="http://roosterbio.blogspot.com/2014/07/best-practices-in-msc-culture-tracking.html"><span style="font-size: 11.0pt;">here</span></a><span style="font-size: 11pt;"> for our popular blog post on PDLs. </span><span style="font-size: 11pt;">Our hUC-MSC exceed this growth rate with a typical expansion of
>20-fold in <b style="mso-bidi-font-weight: normal;">4</b> days </span><span style="font-size: 11pt;">(see
Figure 2A for a representative RoosterBio product donor)</span><span style="font-size: 11pt;">. </span><span style="font-size: 11pt;">We see variability across donors, but donors
are typically harvested at greater than 80,000 cells/cm<sup>2</sup>, after
plating at ~3,000 cells/cm<sup>2</sup>, within 4 days (Figure 2B). So, a vial
of 10M hUC-MSC (</span><span style="font-size: 11pt;">RoosterVial-hUC-10M-XF)</span></span><span style="font-size: 11pt;"><span style="font-family: "arial" , "helvetica" , sans-serif;">
grown over 2 passages has a potential yield of greater than 5B hUC-MSC in 8 to
10 days, leading to significant economic benefits (described below).</span><span style="font-family: "arial" , "helvetica" , sans-serif;"><o:p></o:p></span></span></span></div>
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<span style="color: #444444; font-family: "arial" , "helvetica" , sans-serif;"><b style="mso-bidi-font-weight: normal;"><i style="mso-bidi-font-style: normal;"><span style="font-size: 11pt;"><span style="font-family: "arial" , "helvetica" , sans-serif;">C</span><span style="font-family: "arial" , "helvetica" , sans-serif;">ell surface marker
expression</span></span></i></b><span style="font-family: "arial" , "helvetica" , sans-serif;"><i style="mso-bidi-font-style: normal;"><span style="font-size: 11.0pt;">.<b style="mso-bidi-font-weight: normal;"> </b></span></i><span style="font-size: 11pt;">Flow cytometry
analysis for the ISCT recommended cell surface markers was done for </span><span style="font-size: 11pt;">RoosterVial-hUC</span></span><span style="font-size: 11pt;"><span style="font-family: "arial" , "helvetica" , sans-serif;"> in RoosterNourish™-MSC-XF following a 2
passage expansion. The hUC-MSC population displayed the typical hMSC surface
marker expression: they were low (<5% positive) for the stem and hematopoietic
cell markers, CD14, CD34 and CD45, and >90% positive for hMSC markers CD73,
CD90, CD105, and CD166 (Figure 3A). </span><span style="font-family: "arial" , "helvetica" , sans-serif;"><o:p></o:p></span></span></span></div>
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<span style="color: #444444; font-family: "arial" , "helvetica" , sans-serif;"><b style="mso-bidi-font-weight: normal;"><i style="mso-bidi-font-style: normal;"><span style="font-size: 11pt;">Immunomodulatory function.</span></i></b><span style="font-size: 11pt;"> Immunomodulation is an essential part of the <i style="mso-bidi-font-style: normal;">in vivo</i> therapeutic role(s) of hMSC (Krampera
2013). The immunomodulatory potential of hMSC is assayed<i style="mso-bidi-font-style: normal;"> </i>by activation of indoleamine 2,3-dioxygenase (IDO) activity by the
pro-inflammatory cytokine IFN-γ and measuring kynurenine (kyn), an
immunosuppressive product of the IDO reaction, in the cell culture supernatant.
hUC-MSC were expanded in RoosterNourish™-MSC-XF for 2 passages, harvested, plated,
and treated with IFN-γ. The kynurenine concentration in the cell supernatant
was measured using a spectrophotometric assay. Like other hMSC types, hUC-MSC
showed low basal IDO activity that was inducible by IFN-γ treatment (Figure 3B).
We see variability in induced IDO activity across UC donors (not shown) which
will be a subject of a subsequent blog. Importantly, these data show that RoosterBio’s
hUC-MSC meet the ISCT criterion by demonstrating immunomodulatory activity. <o:p></o:p></span></span></div>
</div>
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<span style="color: #444444; font-family: "arial" , "helvetica" , sans-serif;"><br /></span></div>
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<span style="color: #444444; font-family: "arial" , "helvetica" , sans-serif;"><b style="font-family: arial, helvetica, sans-serif;"><i style="mso-bidi-font-style: normal;"><span style="font-size: 11pt;">Trilineage
differentiation.</span></i></b><i style="font-family: arial, helvetica, sans-serif;"><span style="font-size: 11.0pt;">
</span></i><span style="font-family: "arial" , "helvetica" , sans-serif; font-size: 11pt;">A hallmark characteristic of hMSC and ISCT criterion is differentiation
<i style="mso-bidi-font-style: normal;">in vitro</i> to adipocytes, osteocytes,
and chondrocytes (<span style="color: black; mso-themecolor: text1;">Dominici,
2006; Krampera, 2013</span>). RoosterBio’s hUC-MSCs expanded 2 passages from
the working cell bank in RoosterNourish™-MSC-XF were harvested, plated, and
incubated in Adipogenesis, Osteogenesis, or Chondrogenesis Media. After 18-21
days in culture, staining for adipogenic, osteogenic, and chondrogenic
differentiation potential was performed. As shown in Figure 4, RoosterBio’s hUC-MSC
differentiated to fat, bone and cartilage.</span></span></div>
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<span style="color: #444444; font-family: "arial" , "helvetica" , sans-serif;"><span style="font-family: "arial" , sans-serif;"><span style="font-size: 14.6667px;"><br /></span></span><span style="font-size: 11.0pt;"></span></span></div>
<div class="separator" style="clear: both; text-align: center;">
<span style="color: #444444; font-family: "arial" , "helvetica" , sans-serif; margin-left: 1em; margin-right: 1em;"><a href="https://blogger.googleusercontent.com/img/b/R29vZ2xl/AVvXsEgeLtSUARDgTT_u_BBp3CVY8zt3dqkL9XKQVf3pBbw5uoAs5skvR4XMUZALUlmzvKdxSwoHcwSWUI0yGMit-gohyphenhyphentQvEuQ7vnKnLDcNGCRvLHU8_WvJT6BN_gFizRJZfYtdcvgVdklCToJ_/s1600/Fig+4.jpeg" imageanchor="1" style="margin-left: 1em; margin-right: 1em;"><img border="0" data-original-height="376" data-original-width="996" height="240" src="https://blogger.googleusercontent.com/img/b/R29vZ2xl/AVvXsEgeLtSUARDgTT_u_BBp3CVY8zt3dqkL9XKQVf3pBbw5uoAs5skvR4XMUZALUlmzvKdxSwoHcwSWUI0yGMit-gohyphenhyphentQvEuQ7vnKnLDcNGCRvLHU8_WvJT6BN_gFizRJZfYtdcvgVdklCToJ_/s640/Fig+4.jpeg" width="640" /></a></span></div>
<span style="color: #444444; font-family: "arial" , "helvetica" , sans-serif;"><span style="font-family: inherit;">
</span>
</span><br />
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<span style="color: #444444; font-family: "arial" , "helvetica" , sans-serif;"><br /></span></div>
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<span style="color: #444444; font-family: "arial" , "helvetica" , sans-serif;"><b style="font-family: arial, helvetica, sans-serif;"><i style="mso-bidi-font-style: normal;"><span style="font-size: 11pt;">Potency: Angiogenic
cytokine secretion. </span></i></b><span style="font-size: 11pt;"><span style="font-family: "arial" , "helvetica" , sans-serif;">hMSC achieve their therapeutic effects
by secreting a plethora of biomolecules that influence many biologic processes
(Murphy 2013). As a measure of hUC-MSC potency, we assayed the secretion of the
angiogenic cytokines bFGF, HGF, TIMP 1, TIMP 2, IL-8, and VEGF via a multiplexed
ELISA analysis (Figure 3C). <span style="mso-bidi-font-weight: bold;">When
multiple UC donors are screened we expect hUC-MSC to differ in their cytokine
secretion profile from hAD-MSC and hBM-MSC (some of the cytokines may be higher
or lower, etc). </span></span><span style="font-family: "arial" , "helvetica" , sans-serif;"><o:p></o:p></span></span></span></div>
<div class="MsoNormal" style="text-align: justify;">
<span style="color: #444444; font-family: "arial" , "helvetica" , sans-serif;"><br /></span></div>
<div class="MsoNormal" style="text-align: justify;">
<span style="color: #444444; font-family: "arial" , "helvetica" , sans-serif;"><b style="font-family: arial, helvetica, sans-serif;"><i style="mso-bidi-font-style: normal;"><span style="font-size: 11.0pt;">Potency:
Extracellular vesicle (EV) Production. </span></i></b><span style="font-family: "arial" , "helvetica" , sans-serif;"><span style="font-size: 11pt;">Due to their
similar therapeutic effects to MSCs and potential as a key bioactive agent in
regenerative medicine applications, MSC-derived EVs are being increasingly
investigated in pre-clinical research and as a clinical therapy for a broad
range of indications (<span style="color: black; mso-themecolor: text1;">Elahi,
2019)</span>. However, EV production for these studies is limited by functional
cell number. O</span><span style="font-size: 11pt;">ur</span><span style="font-size: 11pt;"> </span><a href="https://www.roosterbio.com/collections/bioprocess-media"><span style="font-size: 11.0pt;">bioprocess
media systems</span></a></span><span style="font-size: 11pt;"><span style="font-family: "arial" , "helvetica" , sans-serif;"> for cell growth (RoosterNourish™-MSC) and EV
production (<a href="https://www.roosterbio.com/collections/bioprocess-media/products/roostercollect-ev-m2001">RoosterCollect™-EV</a>) are an efficient, scalable system to produce
EVs from hMSC. We have preliminary studies showing robust EV production from
our hUC-MSC (subject of a future blog).</span><span style="font-family: "arial" , "helvetica" , sans-serif;"><o:p></o:p></span></span></span></div>
<div class="MsoNormal">
<span style="color: #444444; font-family: "arial" , "helvetica" , sans-serif;"><br /></span></div>
<div class="MsoNormal" style="text-align: justify;">
<span style="color: #444444; font-family: "arial" , "helvetica" , sans-serif;"><b style="font-family: arial, helvetica, sans-serif;"><i style="mso-bidi-font-style: normal;"><span style="font-size: 11.0pt;">hMSC Tissue origin
differences. </span></i></b><span style="font-family: "arial" , "helvetica" , sans-serif; font-size: 11pt;">As expected, we see variability in
expansion, immunomodulatory activity, differentiation, and angiogenic cytokine
secretion across UC donors (not shown). hUC-MSCs also have tissue-to-tissue
differences when compared to other MSC types. <span style="mso-spacerun: yes;"> </span>Thus, RoosterBio provides at least 3 donors
for each of our hMSC tissue sources allowing customers to screen for the optimal
tissue type and donor for their specific application or target indication. The
relative tissue-specific strengths of hMSC types as well as emerging
application areas will be discussed in subsequent blogs.<span style="mso-bidi-font-weight: bold;"><o:p></o:p></span></span></span></div>
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<span style="color: #444444; font-family: "arial" , "helvetica" , sans-serif;"><br /></span></div>
<div class="MsoNormal" style="text-align: justify;">
<span style="color: #444444; font-family: "arial" , "helvetica" , sans-serif;"><b style="font-family: arial, helvetica, sans-serif;"><i style="mso-bidi-font-style: normal;"><span style="font-size: 11pt;">Productivity Metrics.</span></i></b><span style="font-size: 11pt;">
<span style="font-family: "arial" , "helvetica" , sans-serif;">Due to the very high expansion rates discussed above, the RoosterVial™-hUC,
RoosterNourish™-MSC-XF </span></span><span style="font-family: "arial" , "helvetica" , sans-serif;"><span style="font-size: 11pt;">cell and medium</span><span style="font-size: 11pt;"> </span><span style="font-size: 11pt;">bioprocess system has </span><span style="font-size: 11pt;">unprecedented quantifiable
productivity metrics. The expansion productivity in 2D (<i style="mso-bidi-font-style: normal;">e.g.</i> multi-layer vessels) is ~800 M cells/L of media, an increase
over the already high productivity of our hBM-MSC-XF (200-400 M cells/L). With
our hUC-MSC cell and media system at least 5B cells can be produced in 8-10
days for an investment of <span style="mso-bidi-font-weight: bold;">~$6 to $8K</span></span><span style="font-size: 11pt;">.
</span><span style="font-size: 11pt;">This
productivity increase in also demonstrated in 3D bioreactor and exosome
generation (and will be highlighted in future blog posts).<o:p></o:p></span></span></span></div>
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<span style="color: #444444; font-family: "arial" , "helvetica" , sans-serif;"><br /></span></div>
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<b style="mso-bidi-font-weight: normal;"><span style="color: #444444; font-family: "arial" , "helvetica" , sans-serif; font-size: 11.0pt;">Conclusion<o:p></o:p></span></b></div>
<div class="MsoNormal" style="text-align: justify;">
<span style="color: #444444; font-family: "arial" , "helvetica" , sans-serif; font-size: 11pt;"><br /></span>
<span style="color: #444444; font-family: "arial" , "helvetica" , sans-serif; font-size: 11pt;">RegenMed will source hMSC from multiple tissue types
because each hMSC type will show origin-specific strengths; such as, variations
in expansion capability, immunomodulatory potential, angiogenic cytokine
secretion, tissue-specific differentiation, and in EV production and
characteristics. Therefore, we are introducing XF hUC-MSC (RoosterVial™-hUC-XF)
to expand our portfolio. Having hMSC from multiple tissue sources </span><span style="color: #444444; font-family: "arial" , "helvetica" , sans-serif; font-size: 11pt;">support our customer screening of donor AND tissue source to find their
best hMSC.</span><span style="color: #444444; font-family: "arial" , "helvetica" , sans-serif; font-size: 11pt;"> </span><span style="color: #444444; font-family: "arial" , "helvetica" , sans-serif; font-size: 11pt;">Paired with RoosterBio’s bioprocess media system, </span><span style="color: #444444; font-family: "arial" , "helvetica" , sans-serif; font-size: 11pt;">RoosterNourish™-MSC-XF</span><span style="color: #444444; font-family: "arial" , "helvetica" , sans-serif; font-size: 11pt;">, RoosterVial-hUC reaches new heights in hMSC 2D batch and 3D bioreactor
fed-batch expansion productivity in terms of both M cells/L media and overall
cost, while generating larger volumes (Billions) of hMSCs and EVs.<span style="mso-spacerun: yes;"> </span>Additionally, RoosterBio provides first in
class characterization of hUC-MSC key quality attributes (PDL, identity,
expansion potential) and functional potential (cytokine secretion,
differentiation, immunomodulation). Finally, RoosterBio’s scalable, cGMP
compatible, and ready to implement process recommendations simplify and
accelerate the path through hUC-MSC-based product development to clinical
implementation.</span></div>
<div class="MsoNormal">
<span style="color: #444444; font-family: "arial" , "helvetica" , sans-serif;"><br /></span></div>
<div class="MsoNormal">
<b style="mso-bidi-font-weight: normal;"><span style="color: #444444; font-family: "arial" , "helvetica" , sans-serif; font-size: 11.0pt;">References: <o:p></o:p></span></b></div>
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<span style="color: #444444; font-family: "arial" , "helvetica" , sans-serif;"><span style="font-size: 11.0pt;">Bravery CA, Carmen J, Fong T, Oprea W,
Hoogendoorn KH, Woda J, Burger SR, Rowley JA, Bonyhadi ML, & Van't Hof W
(2013) Potency assay development for cellular therapy products: an ISCT review
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<span style="color: #444444; font-family: "arial" , "helvetica" , sans-serif;"><span style="font-size: 11.0pt;">Carmen J, Burger SR, McCaman M, & Rowley
JA (2012) Developing assays to address identity, potency, purity and safety:
cell characterization in cell therapy process development. <i style="mso-bidi-font-style: normal;">Regenerative Medicine</i> 7(1):85-100. </span><a href="http://www.ncbi.nlm.nih.gov/pubmed/22168500"><span style="font-size: 11.0pt;">http://www.ncbi.nlm.nih.gov/pubmed/22168500</span></a><span style="font-size: 11.0pt;"> <o:p></o:p></span></span></div>
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<span style="color: #444444; font-family: "arial" , "helvetica" , sans-serif;"><span style="font-size: 11.0pt;">Couto
PS, Shatirishvili G, Bersenev A, Verter F. (2019) First decade of clinical
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<span style="color: #444444; font-family: "arial" , "helvetica" , sans-serif;"><span style="font-size: 11.0pt;">Davies JE, Walker JT, Keating A. (2017) <span style="mso-bidi-font-weight: bold;">Concise Review: Wharton's Jelly: The Rich,
but Enigmatic, Source of Mesenchymal Stromal Cells. Stem Cells Transl Med. 6(7):1620-1630.
</span></span><a href="https://www.ncbi.nlm.nih.gov/pubmed/28488282"><span style="font-size: 11.0pt;">https://www.ncbi.nlm.nih.gov/pubmed/28488282</span></a><span style="font-size: 11.0pt;"> <b><o:p></o:p></b></span></span></div>
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<span style="color: #444444; font-family: "arial" , "helvetica" , sans-serif;"><span style="font-size: 11.0pt;">Dominici M, Le Blanc K, Mueller I,
Slaper-Cortenbach I, Marini F, Krause D, Deans R, Keating A, Prockop D, &
Horwitz E (2006) Minimal criteria for defining multipotent mesenchymal stromal
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<span style="color: #444444; font-family: "arial" , "helvetica" , sans-serif;"><span style="font-size: 11.0pt;">Elahi FM, Farwell DG, Nolta JA, Anderson JD
(2019) Concise Review: Preclinical Translation of Exosomes Derived from
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<span style="color: #444444; font-family: "arial" , "helvetica" , sans-serif;"><span style="font-size: 11.0pt;">Krampera M, Galipeau J, Shi Y, Tarte K,
Sensebe L, & MSC Committee of the International Society for Cellular
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<span style="color: #444444; font-family: "arial" , "helvetica" , sans-serif;"><span style="font-size: 11.0pt;">Moll
G, Ankrum JA, Kamhieh-Milz J, Bieback K, Ringdén O, Volk HD, Geissler S, Reinke
P (2019) Intravascular Mesenchymal Stromal/Stem Cell Therapy Product
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<span style="color: #444444; font-family: "arial" , "helvetica" , sans-serif;"><span style="font-size: 11.0pt;">Murphy MB, Moncivais K, & Caplan AI
(2013) Mesenchymal stem cells: environmentally responsive therapeutics for
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(2018) Peak MSC – are we there yet? <i style="mso-bidi-font-style: normal;">Front.
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J, Yu G, Cai M, Lei X, Yang Y, Wang Q, Zhai X (2018) Bibliometric analysis of
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Research & Therapy</i> 9(32). </span><a href="https://www.ncbi.nlm.nih.gov/pubmed/29415771"><span style="font-size: 11pt;">https://www.ncbi.nlm.nih.gov/pubmed/29415771</span></a></span><span style="font-family: "arial" , "helvetica" , sans-serif; font-size: 11pt;"><o:p></o:p></span></span></div>
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<w:LsdException Locked="false" Priority="39" SemiHidden="true"
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<w:LsdException Locked="false" Priority="39" SemiHidden="true"
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<w:LsdException Locked="false" Priority="39" SemiHidden="true"
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<w:LsdException Locked="false" Priority="39" SemiHidden="true"
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<w:LsdException Locked="false" Priority="39" SemiHidden="true"
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<w:LsdException Locked="false" SemiHidden="true" UnhideWhenUsed="true"
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<w:LsdException Locked="false" SemiHidden="true" UnhideWhenUsed="true"
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<w:LsdException Locked="false" SemiHidden="true" UnhideWhenUsed="true"
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<w:LsdException Locked="false" SemiHidden="true" UnhideWhenUsed="true"
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<w:LsdException Locked="false" SemiHidden="true" UnhideWhenUsed="true"
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<w:LsdException Locked="false" SemiHidden="true" UnhideWhenUsed="true"
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<w:LsdException Locked="false" Priority="35" SemiHidden="true"
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<w:LsdException Locked="false" SemiHidden="true" UnhideWhenUsed="true"
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<w:LsdException Locked="false" SemiHidden="true" UnhideWhenUsed="true"
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<w:LsdException Locked="false" SemiHidden="true" UnhideWhenUsed="true"
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<w:LsdException Locked="false" SemiHidden="true" UnhideWhenUsed="true"
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<w:LsdException Locked="false" SemiHidden="true" UnhideWhenUsed="true"
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<w:LsdException Locked="false" SemiHidden="true" UnhideWhenUsed="true"
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<w:LsdException Locked="false" SemiHidden="true" UnhideWhenUsed="true"
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<w:LsdException Locked="false" SemiHidden="true" UnhideWhenUsed="true"
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<w:LsdException Locked="false" SemiHidden="true" UnhideWhenUsed="true"
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<w:LsdException Locked="false" SemiHidden="true" UnhideWhenUsed="true"
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<w:LsdException Locked="false" SemiHidden="true" UnhideWhenUsed="true"
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<w:LsdException Locked="false" SemiHidden="true" UnhideWhenUsed="true"
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<w:LsdException Locked="false" SemiHidden="true" UnhideWhenUsed="true"
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<w:LsdException Locked="false" SemiHidden="true" UnhideWhenUsed="true"
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<w:LsdException Locked="false" SemiHidden="true" UnhideWhenUsed="true"
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<w:LsdException Locked="false" SemiHidden="true" UnhideWhenUsed="true"
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<w:LsdException Locked="false" SemiHidden="true" UnhideWhenUsed="true"
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<w:LsdException Locked="false" SemiHidden="true" UnhideWhenUsed="true"
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<w:LsdException Locked="false" SemiHidden="true" UnhideWhenUsed="true"
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<w:LsdException Locked="false" Priority="10" QFormat="true" Name="Title"/>
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<w:LsdException Locked="false" SemiHidden="true" UnhideWhenUsed="true"
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<w:LsdException Locked="false" Priority="1" SemiHidden="true"
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<w:LsdException Locked="false" SemiHidden="true" UnhideWhenUsed="true"
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<w:LsdException Locked="false" SemiHidden="true" UnhideWhenUsed="true"
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<w:LsdException Locked="false" SemiHidden="true" UnhideWhenUsed="true"
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<w:LsdException Locked="false" SemiHidden="true" UnhideWhenUsed="true"
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<w:LsdException Locked="false" SemiHidden="true" UnhideWhenUsed="true"
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<w:LsdException Locked="false" SemiHidden="true" UnhideWhenUsed="true"
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<w:LsdException Locked="false" SemiHidden="true" UnhideWhenUsed="true"
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<w:LsdException Locked="false" SemiHidden="true" UnhideWhenUsed="true"
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<w:LsdException Locked="false" SemiHidden="true" UnhideWhenUsed="true"
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<w:LsdException Locked="false" SemiHidden="true" UnhideWhenUsed="true"
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<w:LsdException Locked="false" SemiHidden="true" UnhideWhenUsed="true"
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<w:LsdException Locked="false" Priority="22" QFormat="true" Name="Strong"/>
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<w:LsdException Locked="false" SemiHidden="true" UnhideWhenUsed="true"
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<w:LsdException Locked="false" SemiHidden="true" UnhideWhenUsed="true"
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<w:LsdException Locked="false" SemiHidden="true" UnhideWhenUsed="true"
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<w:LsdException Locked="false" SemiHidden="true" UnhideWhenUsed="true"
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<w:LsdException Locked="false" SemiHidden="true" UnhideWhenUsed="true"
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<w:LsdException Locked="false" SemiHidden="true" UnhideWhenUsed="true"
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<w:LsdException Locked="false" SemiHidden="true" UnhideWhenUsed="true"
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<w:LsdException Locked="false" SemiHidden="true" UnhideWhenUsed="true"
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<w:LsdException Locked="false" SemiHidden="true" UnhideWhenUsed="true"
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<w:LsdException Locked="false" SemiHidden="true" UnhideWhenUsed="true"
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<w:LsdException Locked="false" SemiHidden="true" UnhideWhenUsed="true"
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<w:LsdException Locked="false" SemiHidden="true" UnhideWhenUsed="true"
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<w:LsdException Locked="false" SemiHidden="true" UnhideWhenUsed="true"
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<w:LsdException Locked="false" SemiHidden="true" UnhideWhenUsed="true"
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<w:LsdException Locked="false" SemiHidden="true" UnhideWhenUsed="true"
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<w:LsdException Locked="false" SemiHidden="true" UnhideWhenUsed="true"
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<w:LsdException Locked="false" SemiHidden="true" UnhideWhenUsed="true"
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<w:LsdException Locked="false" SemiHidden="true" UnhideWhenUsed="true"
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<w:LsdException Locked="false" SemiHidden="true" UnhideWhenUsed="true"
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<w:LsdException Locked="false" SemiHidden="true" UnhideWhenUsed="true"
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<w:LsdException Locked="false" SemiHidden="true" UnhideWhenUsed="true"
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<w:LsdException Locked="false" SemiHidden="true" UnhideWhenUsed="true"
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<w:LsdException Locked="false" SemiHidden="true" UnhideWhenUsed="true"
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<w:LsdException Locked="false" SemiHidden="true" UnhideWhenUsed="true"
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<w:LsdException Locked="false" SemiHidden="true" UnhideWhenUsed="true"
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<w:LsdException Locked="false" SemiHidden="true" UnhideWhenUsed="true"
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<w:LsdException Locked="false" SemiHidden="true" UnhideWhenUsed="true"
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<w:LsdException Locked="false" SemiHidden="true" UnhideWhenUsed="true"
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<w:LsdException Locked="false" SemiHidden="true" UnhideWhenUsed="true"
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<w:LsdException Locked="false" SemiHidden="true" UnhideWhenUsed="true"
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<w:LsdException Locked="false" SemiHidden="true" UnhideWhenUsed="true"
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<w:LsdException Locked="false" SemiHidden="true" UnhideWhenUsed="true"
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<w:LsdException Locked="false" SemiHidden="true" UnhideWhenUsed="true"
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<w:LsdException Locked="false" SemiHidden="true" UnhideWhenUsed="true"
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<w:LsdException Locked="false" SemiHidden="true" UnhideWhenUsed="true"
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<w:LsdException Locked="false" SemiHidden="true" UnhideWhenUsed="true"
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<w:LsdException Locked="false" SemiHidden="true" UnhideWhenUsed="true"
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<w:LsdException Locked="false" Priority="39" Name="Table Grid"/>
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<w:LsdException Locked="false" Priority="62" Name="Light Grid"/>
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<w:LsdException Locked="false" Priority="65" Name="Medium List 1"/>
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<w:LsdException Locked="false" Priority="72" Name="Colorful List"/>
<w:LsdException Locked="false" Priority="73" Name="Colorful Grid"/>
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<w:LsdException Locked="false" Priority="65" Name="Medium List 1 Accent 1"/>
<w:LsdException Locked="false" SemiHidden="true" Name="Revision"/>
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<w:LsdException Locked="false" Priority="30" QFormat="true"
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<w:LsdException Locked="false" Priority="72" Name="Colorful List Accent 1"/>
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<w:LsdException Locked="false" Priority="60" Name="Light Shading Accent 6"/>
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<w:LsdException Locked="false" Priority="19" QFormat="true"
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<w:LsdException Locked="false" Priority="21" QFormat="true"
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<w:LsdException Locked="false" Priority="31" QFormat="true"
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<w:LsdException Locked="false" Priority="32" QFormat="true"
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<w:LsdException Locked="false" Priority="33" QFormat="true" Name="Book Title"/>
<w:LsdException Locked="false" Priority="37" SemiHidden="true"
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<w:LsdException Locked="false" Priority="39" SemiHidden="true"
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<w:LsdException Locked="false" Priority="41" Name="Plain Table 1"/>
<w:LsdException Locked="false" Priority="42" Name="Plain Table 2"/>
<w:LsdException Locked="false" Priority="43" Name="Plain Table 3"/>
<w:LsdException Locked="false" Priority="44" Name="Plain Table 4"/>
<w:LsdException Locked="false" Priority="45" Name="Plain Table 5"/>
<w:LsdException Locked="false" Priority="40" Name="Grid Table Light"/>
<w:LsdException Locked="false" Priority="46" Name="Grid Table 1 Light"/>
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<w:LsdException Locked="false" Priority="48" Name="Grid Table 3"/>
<w:LsdException Locked="false" Priority="49" Name="Grid Table 4"/>
<w:LsdException Locked="false" Priority="50" Name="Grid Table 5 Dark"/>
<w:LsdException Locked="false" Priority="51" Name="Grid Table 6 Colorful"/>
<w:LsdException Locked="false" Priority="52" Name="Grid Table 7 Colorful"/>
<w:LsdException Locked="false" Priority="46"
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<w:LsdException Locked="false" Priority="47" Name="Grid Table 2 Accent 1"/>
<w:LsdException Locked="false" Priority="48" Name="Grid Table 3 Accent 1"/>
<w:LsdException Locked="false" Priority="49" Name="Grid Table 4 Accent 1"/>
<w:LsdException Locked="false" Priority="50" Name="Grid Table 5 Dark Accent 1"/>
<w:LsdException Locked="false" Priority="51"
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<w:LsdException Locked="false" Priority="52"
Name="Grid Table 7 Colorful Accent 1"/>
<w:LsdException Locked="false" Priority="46"
Name="Grid Table 1 Light Accent 2"/>
<w:LsdException Locked="false" Priority="47" Name="Grid Table 2 Accent 2"/>
<w:LsdException Locked="false" Priority="48" Name="Grid Table 3 Accent 2"/>
<w:LsdException Locked="false" Priority="49" Name="Grid Table 4 Accent 2"/>
<w:LsdException Locked="false" Priority="50" Name="Grid Table 5 Dark Accent 2"/>
<w:LsdException Locked="false" Priority="51"
Name="Grid Table 6 Colorful Accent 2"/>
<w:LsdException Locked="false" Priority="52"
Name="Grid Table 7 Colorful Accent 2"/>
<w:LsdException Locked="false" Priority="46"
Name="Grid Table 1 Light Accent 3"/>
<w:LsdException Locked="false" Priority="47" Name="Grid Table 2 Accent 3"/>
<w:LsdException Locked="false" Priority="48" Name="Grid Table 3 Accent 3"/>
<w:LsdException Locked="false" Priority="49" Name="Grid Table 4 Accent 3"/>
<w:LsdException Locked="false" Priority="50" Name="Grid Table 5 Dark Accent 3"/>
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Sandy Applebeehttp://www.blogger.com/profile/01351576729478919332noreply@blogger.com0tag:blogger.com,1999:blog-6181073719515632715.post-33206307265651643852019-08-16T16:59:00.001-04:002019-09-11T15:19:19.155-04:00Behind Our Best Places to Work Award: Life Inside The Roost<div dir="ltr" style="line-height: 1.2; margin-bottom: 0pt; margin-top: 0pt; text-align: center;">
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<span style="font-family: inherit;">We’re thrilled to share that we’ve been named a Frederick County Best Places to Work! This award is especially meaningful to us because our company was built with culture in mind, from our workspaces, our employee benefits, to the way we interact with our customers. <o:p></o:p></span></div>
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<b><span style="font-family: inherit; font-size: large;">What Makes RoosterBio a “Best Place to Work”</span></b></div>
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<span style="font-family: inherit;">The Frederick County </span><a href="http://www.frederickbestplacestowork.com/" style="color: #954f72; font-family: inherit;">Best Places to Work</a><span style="font-family: inherit;"> is an annual campaign that showcases a company’s best practices to create an amazing place to work. The survey includes several open-ended questions centered around key components that help determine a winner. While an emphasis is placed on average median salaries and voluntary turnover rates, we wanted to share some of the key parts of our company’s culture that address other aspects of the award criteria. At RoosterBio, we’re serious about the work we do but we try not to take ourselves too seriously, and we have a little fun along the way!</span></div>
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<span style="font-family: inherit;">Take a peek inside life at The Roost.</span><span style="font-family: "calibri" , sans-serif;"><o:p></o:p></span><br />
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<a href="https://blogger.googleusercontent.com/img/b/R29vZ2xl/AVvXsEhRE7X50IOLJSRSlCSu2zw-aDFjpmszvjEsSL6cuW6nvEIRAtcozXEKYuplL8-RtT4Ur4eaUT4s0xmD6A_M98aLuPdlmT7Tw9-o9-4ZNnkIsn7l1SfeIx2jf7YWtDxNZakxuO2klaChc_iA/s1600/BPicture1.png" imageanchor="1" style="margin-left: 1em; margin-right: 1em;"><img border="0" data-original-height="428" data-original-width="643" height="424" src="https://blogger.googleusercontent.com/img/b/R29vZ2xl/AVvXsEhRE7X50IOLJSRSlCSu2zw-aDFjpmszvjEsSL6cuW6nvEIRAtcozXEKYuplL8-RtT4Ur4eaUT4s0xmD6A_M98aLuPdlmT7Tw9-o9-4ZNnkIsn7l1SfeIx2jf7YWtDxNZakxuO2klaChc_iA/s640/BPicture1.png" width="640" /></a></div>
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<b><span style="font-family: inherit;"><span style="font-size: large;">Attracting the Best Talent</span><o:p></o:p></span></b></div>
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<a href="https://blogger.googleusercontent.com/img/b/R29vZ2xl/AVvXsEiSth-gmJQWtXVWrOU7W5udKUE_PaqOW88xckPgD8Yojuxzc1fMUqMz4LDyBxUtfxBW9sKFMv-35znw25Sul-9dcjC2eMT2E9ev5Cogy2YPC5z92a247bsftml1sEfzeAhw_CNsl54Tunvl/s1600/BPicture2.png" imageanchor="1" style="clear: right; float: right; margin-bottom: 1em; margin-left: 1em;"><img border="0" data-original-height="422" data-original-width="316" height="320" src="https://blogger.googleusercontent.com/img/b/R29vZ2xl/AVvXsEiSth-gmJQWtXVWrOU7W5udKUE_PaqOW88xckPgD8Yojuxzc1fMUqMz4LDyBxUtfxBW9sKFMv-35znw25Sul-9dcjC2eMT2E9ev5Cogy2YPC5z92a247bsftml1sEfzeAhw_CNsl54Tunvl/s320/BPicture2.png" width="233" /></a><b><span style="font-family: inherit;"><br /></span></b></div>
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<span style="font-family: inherit;">RoosterBio is a pioneer at the forefront of a groundbreaking regenerative medicine industry with cutting-edge technology and science that attracts the brightest people from a variety of backgrounds. Employee equity in the company where everyone is a shareholder, a collaborative work environment with an open floor plan, community meeting spaces as well as a modern, open and airy laboratory space to work in - are all perks of being a Rooster.</span></div>
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<span style="font-family: inherit;">We regularly host higher education students from across the country who are looking to further their experience in the lab, from Harvard University to the University of Virginia this year, this kind of partnership helps spread the word among academia about the kind of innovative work RoosterBio is doing. </span><span style="font-family: "calibri" , sans-serif;"><o:p></o:p></span></div>
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<span style="font-family: inherit;">One of the partners we work with is the National Institute for Innovation in Manufacturing Biopharmaceuticals (NIIMBL) and the National Society of Black Engineers. This NIIMBL experience includes a tour and panel discussion to give students enrolled at historically black colleges and universities (HBCUs) an understanding of the kinds of careers available within biopharmaceutical manufacturing.</span></div>
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<a href="https://blogger.googleusercontent.com/img/b/R29vZ2xl/AVvXsEilOqE_Igs83237PPyFBFPOmAkxSQcaSrEuZDpmZ3iXIDoMNNe7_62o34pB3K0luyKHSC0jkDqL_5_0yCUDgeTZv36s1K9blP_a3Khe4VuQyQsr5hvw9VystUSyFjej_9b3TT0mAqNt09gL/s1600/BPicture3.png" imageanchor="1" style="margin-left: 1em; margin-right: 1em;"><img border="0" data-original-height="368" data-original-width="491" height="478" src="https://blogger.googleusercontent.com/img/b/R29vZ2xl/AVvXsEilOqE_Igs83237PPyFBFPOmAkxSQcaSrEuZDpmZ3iXIDoMNNe7_62o34pB3K0luyKHSC0jkDqL_5_0yCUDgeTZv36s1K9blP_a3Khe4VuQyQsr5hvw9VystUSyFjej_9b3TT0mAqNt09gL/s640/BPicture3.png" width="640" /></a></div>
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<b><span style="font-family: inherit;"><span style="font-size: large;">Retaining a Strong Workforce</span><o:p></o:p></span></b></div>
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<span style="font-family: inherit;">Retaining our workforce is so critical that Building and Nurturing Company Culture is one of RoosterBio’s corporate and strategic objectives. It’s not just something on a piece of paper, but something we nurture and invest in every day.</span><span style="font-family: "calibri" , sans-serif;"><o:p></o:p></span></div>
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<span style="font-family: inherit;">The leadership team at RoosterBio believes in providing ongoing access to training and promotes transparency so much so that there is no question off-limits during our quarterly “All-Hands” Meetings. Our employees are highly engaged as measured by ongoing and anonymous surveys.</span></div>
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<a href="https://blogger.googleusercontent.com/img/b/R29vZ2xl/AVvXsEgbv0UhwlFQRa-N_pj_OnSFTa2C9fXITGeUFN9JdjEXk7DXgNbPb5AfMSFjnBd57fGFiAApuijwE7hMpj0BRKYfJR3ccBWziyda8WUyba7Q_E1D7yk6v9ffQPo8OeIOp4V1dV5mRggftg35/s1600/BPicture4.png" imageanchor="1" style="margin-left: 1em; margin-right: 1em;"><img border="0" data-original-height="375" data-original-width="497" height="482" src="https://blogger.googleusercontent.com/img/b/R29vZ2xl/AVvXsEgbv0UhwlFQRa-N_pj_OnSFTa2C9fXITGeUFN9JdjEXk7DXgNbPb5AfMSFjnBd57fGFiAApuijwE7hMpj0BRKYfJR3ccBWziyda8WUyba7Q_E1D7yk6v9ffQPo8OeIOp4V1dV5mRggftg35/s640/BPicture4.png" width="640" /></a></div>
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<b><span style="font-size: large;"><b><span style="font-family: inherit; font-size: large;"><br /></span></b></span></b></div>
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<a href="https://blogger.googleusercontent.com/img/b/R29vZ2xl/AVvXsEgZnfGFWoGBAOgOkJmQvdNViWVY1e-t1g4e3SXPQkePr6cqnxDs2mfBqFyV0G_nKg7G6_b1sqk8mi1xdIQw0SuFpYA2Pgz_NHXfBlBQcM_3xVwJp7GQH7knNOMWEjRjVDX-x-xC6cBnqj3u/s1600/BPicture5.png" imageanchor="1" style="clear: right; float: right; margin-bottom: 1em; margin-left: 1em;"><img border="0" data-original-height="364" data-original-width="360" height="320" src="https://blogger.googleusercontent.com/img/b/R29vZ2xl/AVvXsEgZnfGFWoGBAOgOkJmQvdNViWVY1e-t1g4e3SXPQkePr6cqnxDs2mfBqFyV0G_nKg7G6_b1sqk8mi1xdIQw0SuFpYA2Pgz_NHXfBlBQcM_3xVwJp7GQH7knNOMWEjRjVDX-x-xC6cBnqj3u/s320/BPicture5.png" width="313" /></a><b><span style="font-size: large;"><b><span style="font-family: inherit; font-size: large;">Cool Perks</span></b></span></b></div>
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<span style="font-family: inherit;">RoosterBio’s workplace was built with Company Culture in mind, from glass doors on offices to promote openness, a collaborative community room to a fully-stocked kitchen that serves as the gathering spot for social events. Flexible work schedules, unlimited paid time off, a robust benefits package, access to things like financial advisors, travel planning discounts, discounted gym memberships, discounted child care and car buying discounts. In addition to 10 paid company holidays, and a floating holiday, we often shut down before holidays that isn’t a part of the communicated holiday calendar.</span></div>
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<b><span style="font-family: inherit; font-size: large;">Promoting Fun at Work</span></b></div>
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<span style="font-family: inherit;">Fun at work matters at RoosterBio because people work harder, stay longer and take better care of the organization when they’re not stressed out. The RoosterBio Culture Club plans events in five main areas: Onsite Fun, Offsite Fun, Volunteering/ Community Outreach, Employee Recognition and Health/Wellness with the mission to <i>Make Work Awesome</i>. We’ve celebrated everything from Take Your Child to Work Day and major holidays to things like International Haiku Day and National Scrabble Day. We recently held the 1st Annual RoosterGames at the RoosterBio Company Picnic where we put our teamwork and perseverance to the ultimate test! To relieve stress at the end of the day, you might hear the faint sound of a ping-pong ball in the distance or catch a group of Roosters at an after-hours happy hour.</span></div>
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<b><span style="font-family: inherit; font-size: large;">Celebrating Success Together</span></b></div>
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<span style="font-family: inherit;">Individual efforts are important but it’s team work that makes the dream work within The Roost. The Golden Rooster awards are for those employees who exemplify company values. These awards are nominated by fellow team members and given quarterly at “All Hands” meetings. Bonuses, additional company equity and work-sponsored celebrations are all ways that success is shared.</span><span style="font-family: "calibri" , sans-serif;"><o:p></o:p></span></div>
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<b><span style="font-family: inherit;"><span style="font-size: large;">Not Just Culture Cluck</span><o:p></o:p></span></b></div>
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<span style="font-family: inherit;">We are proud of what we’ve achieved together and it’s a great time to be a Rooster with even more exciting opportunities ahead. Check out our Introducing Our #Roosters campaign on <a href="https://www.linkedin.com/search/results/all/?keywords=roosterbio&origin=GLOBAL_SEARCH_HEADER" style="color: #954f72;">LinkedIn</a> and <a href="https://www.facebook.com/RoosterBio2013/" style="color: #954f72;">Facebook</a> for testimonials from almost half of the company on what they decided to join RoosterBio. <o:p></o:p></span></div>
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<span style="font-family: inherit;"><a href="https://www.roosterbio.com/pages/join-our-team">Check out our open positions</a> as we expand our presence across the globe and be a part of this exciting journey.</span><span style="font-family: "calibri" , sans-serif;"><o:p></o:p></span></div>
Sandy Applebeehttp://www.blogger.com/profile/01351576729478919332noreply@blogger.com0tag:blogger.com,1999:blog-6181073719515632715.post-65900184698351251262019-07-15T16:58:00.003-04:002019-07-18T09:26:14.292-04:00Advances in Clinical Translation and Scale-up of MSCs and Extracellular Vesicles at ISCT 2019<div class="MsoNormal" style="margin: 0in 0in 0.0001pt; text-align: justify;">
<i><span style="font-family: inherit;">Authored by Katrina Adlerz, Ph.D., Scientist, Analytics, Product & Process Development<o:p></o:p></span></i></div>
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<i><span style="font-family: inherit;">& Josephine Lembong, Ph.D., Scientist, Analytics, Product & Process Development<o:p></o:p></span></i></div>
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<a href="https://blogger.googleusercontent.com/img/b/R29vZ2xl/AVvXsEibDBDmwvIT_HNubATberWEPY4MmycczHwSEhA70d4aGFzvLj2KEoVpQMj5bn86x9ly8A0Lp-WlqICTfQWOOJ43Bb-OjcWskYN9sfnQPFzBSVNPUTQxRbPAQc4ZdYxz8Jbk0g3gnnm7zc4R/s1600/ISCT2019-Globe.jpg" imageanchor="1" style="clear: right; float: right; margin-bottom: 1em; margin-left: 1em;"><img border="0" data-original-height="545" data-original-width="1200" height="145" src="https://blogger.googleusercontent.com/img/b/R29vZ2xl/AVvXsEibDBDmwvIT_HNubATberWEPY4MmycczHwSEhA70d4aGFzvLj2KEoVpQMj5bn86x9ly8A0Lp-WlqICTfQWOOJ43Bb-OjcWskYN9sfnQPFzBSVNPUTQxRbPAQc4ZdYxz8Jbk0g3gnnm7zc4R/s320/ISCT2019-Globe.jpg" width="320" /></a></div>
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<span style="font-family: inherit;">The <span style="color: #954f72;"><a href="http://www.isct2019.com/" style="color: #954f72;">2019 International Society for Cell and Gene Therapy (ISCT) Annual Meeting</a> </span>brought clinicians, regulators, and industry to Melbourne, Australia to collaborate and share progress in the rapidly developing field of Cell & Gene Therapy. Mesenchymal Stem/Stromal Cells (MSCs) were a major focus of the conference with an entire preconference workshop devoted to the <span style="color: #954f72;"><a href="http://www.isct2019.com/msc-workshop-state-of-the-art-in-mscs/#1549498463814-6282c183-ab0d" style="color: #954f72;">Global Clinical Trial Landscape of MSCs</a> </span>as well as multiple sessions and keynotes throughout the conference. RoosterBio was active in the technical sessions and presented new technology on the industrial scale up of both MSCs and MSC-derived extracellular vesicles, as well as announcing a partnership with <span style="color: #954f72;"><a href="https://www.prweb.com/releases/roosterbio_and_tissue_regeneration_therapeutics_collaborate_to_make_human_umbilical_cord_derived_msc_bioprocess_systems_broadly_available/prweb16341107.htm" style="color: #954f72;">Tissue Regeneration Therapeutics</a> </span>to <a href="https://www.prweb.com/releases/roosterbio_and_tissue_regeneration_therapeutics_collaborate_to_make_human_umbilical_cord_derived_msc_bioprocess_systems_broadly_available/prweb16341107.htm">make umbilical cord -derived MSCs broadly available</a>. Other key themes at this year’s meeting were: addressing global regulatory compliance, scalability, reducing cost of goods, exciting developments in exosomes/extracellular vesicles and recent advances in many different cell and gene therapy strategies.<o:p></o:p></span></div>
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<b><span style="font-family: inherit;">MSC Progress in Clinical Trials, Manufacturing, and Comparability<o:p></o:p></span></b></div>
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<span style="font-family: inherit;">Progress in clinical trials across the globe was the focus of a full day pre-conference workshop. Dr. Robert Mays of Athersys provided an update on their stromal cell product Multistem as they move into <span style="color: #954f72;"><a href="https://www.athersys.com/clinical-trials/ischemic-stroke/default.aspx" style="color: #954f72;">Phase III clinical trials for ischemic stroke patients</a> </span>and continue to investigate the treatment’s mechanism of action with immune regulation possibly being one key piece. Dr. Yufang Shi echoed this, highlighting work to pre-condition MSCs to bolster immune responses. Dr. Eleuterio Lombardo of Takeda discussed the development of <a href="https://www.drugdevelopment-technology.com/projects/alofisel-darvadstrocel-treatment-complex-perianal-fistulas-crohns-disease/" style="color: #954f72;">Alofisel, an allogeneic MSC product</a> that has been approved in Europe for perianal fistulas in Crohn’s disease. Some challenges in clinical trials were highlight<table cellpadding="0" cellspacing="0" class="tr-caption-container" style="float: right; margin-left: 1em; text-align: right;"><tbody>
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<tr><td class="tr-caption" style="text-align: center;">Photo courtesy http://www.isct2019.com/photo-gallery/</td></tr>
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ed, such as inconsistent responses between pre-clinical and clinical studies. Dr. Lombardo pointed out an often-stated public opinion that successes in animal studies are often attributed to the use of “fresher” cells, and that these results may not be repeatable in human clinical trials because clinical trials often use cells that have been expanded and cryopreserved. Dr. Lombardo presented his review of the literature which did not support this opinion. Instead, he found that many studies did not state whether MSCs were cryopreserved or fresh, and surprisingly, it was also very rare that studies reported the <a href="http://roosterbio.blogspot.com/2014/07/best-practices-in-msc-culture-tracking.html?utm_content=94850981&utm_medium=social&utm_source=linkedin&hss_channel=lcp-5092232#more" style="color: #954f72;">Population Doubling Level (PDL)</a>. Furthermore, in his recent study where fresh and cryopreserved cells were </span><br />
<a name='more'></a><span style="font-family: inherit;">directly compared in a pneumosepsis model, he observed that fresh cells did not offer a significant advantage [<a href="https://doi.org/10.1002/sctm.18-0260" style="color: #954f72;">1</a>]. <span style="color: #954f72;"><a href="http://www.verypowerfulbiology.com/" style="color: #954f72;">Dr. John Davies of Tissue Regeneration Therapeutics</a> </span>presented on umbilical cord MSCs as a platform for <a href="http://www.verypowerfulbiology.com/news/TRT_StemCellsPaper_PressRelease_08152016.pdf" style="color: #954f72;">gene therapy</a>. RoosterBio’s founder and Chief Product Officer Dr. Jon A. Rowley spoke during the pre-conference MSC workshop on RoosterBio’s innovative strategy of creating standardized and scalable cell and media systems for the Regenerative Medicine Industry, and highlighted paths for radically shortening product development timelines and shortening path to the clinic (you can download the presentation <a href="https://info.roosterbio.com/roosterbio-isct2019-follow-up-0-0-2">here</a><span id="goog_1934116114"></span><span id="goog_1934116115"></span><a href="https://www.blogger.com/"></a>). Dr. Josephine Lembong of RoosterBio also presented a poster on comparability of 2D flasks and 3D bioreactor expanded hMSCs, with comparability of the two systems across growth rates, PDLs, flow markers, multi lineage differentiation, angiogenic cytokine secrection as well as inducible immunomodulatory functions of the cells (download her poster <a href="https://info.roosterbio.com/roosterbio-isct2019-follow-up-0-0-2">here</a>)<o:p></o:p></span></div>
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<b><span style="font-family: inherit;">Addressing a Global Market<o:p></o:p></span></b></div>
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<span style="font-family: inherit;"><span style="font-family: inherit;">The challenge of harmonizing global regulations and manufacturing cell therapy products for global compliance was a common thread during the conference. Discussions centered around the challenge of understanding and complying with different countries’ regulations related to donor screening, raw materials, manufacturing, and clinical trial design, with calls for more uniformity across regions. It was noted that each geographical region has its own regulatory stringency, and that accelerated regulatory frameworks present in certain regions play a role in </span>the timeline for a product’s market approval. For example, Dr. <a href="https://www.hemacare.com/quality-standards/" style="color: #954f72;">Dominic Clarke from HemaCare</a> discussed some differences in donor screening requirements between the US and EU. Marlin Frechette, Director of QA/RA at Irvine Scientific discussed using risk assessments early in the design process to keep manufacturing consistent for a product being produced and distributed across the world. Specifically, she discussed avoiding development of a product that might not meet global regulatory requirements and including this as an early design requirement. Dr. Kilian Kelly from <a href="https://www.cynata.com/" style="color: #954f72;">Cynata</a> talked about the logistical challenges associated with having one global manufacturing facility and multiple regional depots. For example, there are different requirements for donor preference and batch release based on the region. Tony Lee from Medipost emphasized the importance of clinical development strategy and clinical trial study design which may be different depending on the market. Their <span style="color: #954f72;"><a href="http://www.medi-post.com/front/eng/stemcell/cartistem.do" style="color: #954f72;">CartiStem product</a> </span>for osteoarthritis was successful in this regard with market approval in Korea and clinical trial approval by the US FDA. <o:p></o:p></span></div>
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<b><span style="font-family: inherit;">New Advances in EVs <o:p></o:p></span></b></div>
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<span style="font-family: inherit;">Extracellular Vesicles were highlighted in multiple sessions with presentations ranging from preclinical research to clinical translation. Dr. Sai-Kiang Lim chaired a session titled “Challenges in Translating MSC EVs into the Clinic” where the challenges of EV characterization and manufacturing were discussed. <span style="color: #954f72;"><a href="https://scholar.google.com/citations?user=rksfUdkAAAAJ&hl=de" style="color: #954f72;">Dr. Bernd Giebel of Essen University Hospital</a> </span>presented on the development of a Mixed Lymphocyte Assay to better predict potency in animal models. Dr. Elizabeth Shpall of MD Anderson presented on progress moving towards clinical trials for pancreatic cancer with MSC EVs. Posters related to EVs ranged from demonstrating therapeutic effects in animal models to downstream processing. Dr. Katrina Adlerz from RoosterBio presented posters related to RoosterBio’s cell and media systems for rapidly generating significant volumes of MSC-EVs for both research and product development, strategies for increasing EV yield, quality characteristics of MSC-EVs, as well as manufacturing EVs in bioreactor systems with 2D comparability (find a link to the poster <a href="https://info.roosterbio.com/roosterbio-isct2019-follow-up-0-0-2">here</a><span id="goog_1934116122"></span><span id="goog_1934116123"></span><a href="https://www.blogger.com/"></a>). <o:p></o:p></span></div>
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<b><span style="font-family: inherit;">Tackling Business Model Challenges<o:p></o:p></span></b></div>
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<span style="font-family: inherit;">Creating a more sustainable Regenerative Medicine business model by lowering Cost of Goods (COGs) of therapeutic products via manufacturing efficiencies was also a popular topic throughout the Commercialization Tracks. Dr. Jon Rowley presented a talk around reducing COGs by enhancing media productivity to help enable patients’ access to affordable cell and gene therapies. The take home message was that cell culture media is often the largest cost driver of cell therapy product<table cellpadding="0" cellspacing="0" class="tr-caption-container" style="float: right; margin-left: 1em; text-align: right;"><tbody>
<tr><td style="text-align: center;"><a href="https://blogger.googleusercontent.com/img/b/R29vZ2xl/AVvXsEjoKgt4AJLX2h2Y2do0e7fH6zgERH_w8a96OYoyoVdzFn2jjnFRzjX-FUvFlymhUM7w7LfrIagiXniYTKNe0wX1hyphenhyphenITFgmYIt9gwMEUfiuofCGSdSvzkNSpPmW6EYPzuBFtCKyu4AE_ucAE/s1600/20190530-0108.jpg" imageanchor="1" style="clear: right; margin-bottom: 1em; margin-left: auto; margin-right: auto;"><img border="0" data-original-height="533" data-original-width="800" height="266" src="https://blogger.googleusercontent.com/img/b/R29vZ2xl/AVvXsEjoKgt4AJLX2h2Y2do0e7fH6zgERH_w8a96OYoyoVdzFn2jjnFRzjX-FUvFlymhUM7w7LfrIagiXniYTKNe0wX1hyphenhyphenITFgmYIt9gwMEUfiuofCGSdSvzkNSpPmW6EYPzuBFtCKyu4AE_ucAE/s400/20190530-0108.jpg" width="400" /></a></td></tr>
<tr><td class="tr-caption" style="text-align: center;">Photo Courtesy http://www.isct2019.com/photo-gallery/</td></tr>
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s, but that it is also critical to realize the “hidden costs” that are associated with media – as this concept is applicable to everyone working in the cell therapy field. Automation was also presented as a key to COGs reduction. <a href="https://www.at-vio.com/" style="color: #954f72;">ATVIO Biotech, an Orgenesis business</a>, showcased an innovatively designed automated, fully closed bioreactor product called the ADVA, stirred tank CART cell manufacturing platform, with integrated wash & volume reduction capability that could drive overall cost down by >50% - as well as potentially making these therapies available bedside in the clinic.<o:p></o:p></span></div>
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<span style="font-family: inherit;">As the cell therapy field continues to address scalability, new technologies were presented. Various bioreactor platforms were highlighted: PBS Biotech with their vertical-wheel technology, Terumo with their hollow-fiber technology, and Pall’s stirred tank bioreactors, among many others. Downstream processing platforms are equally critical. For example, the Gibco CTS Rotea Counterflow Centrifugation System was highlighted, as well as Terumo’s automated fill and finish system. These technologies were developed for a range of applications; therefore it is always important to go through the exercise to <a href="http://roosterbio.blogspot.com/2018/12/building-effective-multi-year-process.html" style="color: #954f72;">evaluate and determine which technology works for your intended application and scale</a>. <o:p></o:p></span></div>
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<b><span style="font-family: inherit;">Recent Advances in Other Cell Therapies<o:p></o:p></span></b></div>
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<span style="font-family: inherit;">In addition to MSCs, the conference highlighted recent advances in exciting cell therapies focusing on immune oncology, tissue engineering, and regenerative medicine. Dr. Saar Gill from the University of Pennsylvania talked about his approach with CAR T-cell therapy to overcome the absence of cancer specific surface markers [<a href="https://doi.org/10.1016/j.cell.2018.05.013" style="color: #954f72;">2</a>]. Development of iPSC-derived MSC therapies were also highlighted with an emphasis on the capability to achieve large-scale manufacturing from one single donor. Igor Sluvkin, co-founder of Cynata Therapeutics, spoke about their manufacturing capability of producing 19M clinical doses of their iPSC-derived <span style="color: #954f72;"><a href="https://www.cynata.com/about-cymerus" style="color: #954f72;">Cymerus™</a> </span>MSC product from one single donor. Dr. John Rasko from The University of Sydney School of Medicine presented the results of the successful Phase I Cymerus clinical trial for acute GvHD. This is a promising approach to overcome supply limitation, reduce COGs, and also limit donor to donor variability, which seems to be a large issue being tackled by product developers utilizing adult MSC sources. <b><o:p></o:p></b></span></div>
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<span style="font-family: inherit;">Overall, the excitement in the cell and gene therapy field, and really all of Regenerative Medicine, continues to build and we look forward to next year’s meeting in Paris!<o:p></o:p></span></div>
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<span style="font-family: inherit;"><i>References:</i><o:p></o:p></span></div>
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<span style="font-family: inherit;">1. Perlee et al., 2019. Human Adipose‐Derived Mesenchymal Stem Cells Modify Lung Immunity and Improve Antibacterial Defense in Pneumosepsis Caused by Klebsiella pneumoniae. <i>Stem Cells Transl Med</i>. doi: 10.1002/sctm.18-0260<o:p></o:p></span></div>
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<span style="font-family: inherit;">2. Kim et al., 2018. Genetic Inactivation of CD33 in Hematopoietic Stem Cells to Enable CAR T Cell Immunotherapy for Acute Myeloid Leukemia. <i>Cell</i>173(6):1439-1453.e19. doi: 10.1016/j.cell.2018.05.013<o:p></o:p></span></div>
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Sandy Applebeehttp://www.blogger.com/profile/01351576729478919332noreply@blogger.com0tag:blogger.com,1999:blog-6181073719515632715.post-19296980264215157702019-06-11T16:07:00.000-04:002019-06-11T16:38:59.935-04:00Cells as Bioinks for 3D Bioprinting<i>Authored by Mayasari Lim, PhD, Regional Account Manager, West Coast, RoosterBio</i><br />
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<b>Bioprinting overview</b><br />
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The field of 3D bioprinting has exploded in recent years largely due to the advances in additive manufacturing technology along with progress in material chemistry and tissue engineering techniques. The key ingredients that make up the complex bioprinted structures are comprised of hydrogel-based biomaterial/s often coined as ‘bioinks’ and a cellular component that serves as building blocks to create a 3D printed biological tissue. Clearly, the choice of the cell source, proteins and other biological ingredients depends largely on the desired final application. For the purpose of this blog, we will only focus on the desire to create bioprinted tissue for clinical translation. <br />
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<b>Which cell should I use?</b><br />
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Cells for clinical use can be derived from the patient (autologous) or a donor (allogeneic). In many tissue engineering applications, stem cells are used due to their properties in self-renewal and differentiation. Adult stem cells, currently being the most clinically viable solution, include hematopoietic, mesenchymal, neural and epithelial. While hematopoietic stem cell transplantation is widespread, it has limited utility in tissue engineering applications due to its limited differentiation capabilities primarily toward blood lineages. Neural stem cells are most effective in neural regeneration but the limited source makes it very challenging to become clinically relevant. Mesenchymal stem/stromal cells (MSCs), on the other hand, has a significant advantage due to its tri-lineage differentiation ability and immunomodulatory functions thus it has been widely used in various therapeutic indications including brain trauma, graft-versus-host disease and cardiovascular disease. Moreover, MSCs have already demonstrated clinical safety in > 800 clinical trials treating over 30,000 patients to-date.<br />
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<b>How many cells do I need?</b><br />
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<a href="https://www.blogger.com/blogger.g?blogID=6181073719515632715" imageanchor="1" style="clear: right; float: right; margin-bottom: 1em; margin-left: 1em;"></a>In order to print a 3D tissue, we need a significant number of cells seeded at a relatively high density to achieve full tissue mimicry. Exactly how many cells would one require? Let us take a look at a simple example of the knee meniscus. The figure below (left) illustrates a 3D model of an adult meniscus with rough dimensions of 3.5 cm in diameter and 5.3 mm in height. If we were to print this structure at 30% infill, it would require a total volume of ~2.5 mL of bioink. Several studies in bioprinting cartilage tissues have reported that cells would need to be seeded at high densities, a minimum of 10-25 million cells/mL in order to form cartilage in vivo[1, 2]. Thus, in this print, we would require roughly 62.5 million MSCs for a single print. For an intervertebral disc with a diameter of 4 cm and height of 10 mm (Figure on the right), the total volume of bioink required to perform a print would be 4 mL thus the total number of cells required would be 100 million for a single print. These two examples serve to illustrate the number of cells required to bioprint a simple 3D tissue. For larger tissues or organs, one can imagine that you will need a significantly higher number of cells, in the orders to 10 billion or more, to achieve desirable cell distribution through the tissue.<br />
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Traditional methods of expanding MSCs in the lab using 2D tissue culture flasks will require significant amount of time (weeks) and labor to obtain enough cell numbers given that most commercially available MSC vials are sold in 0.5-1 million cells/vial. Fortunately, RoosterBio has developed a ready-to-print cell vial (<a href="https://www.roosterbio.com/collections/high-volume-hmscs/products/roosterrtp-hbm-50m-msc-006">RoosterRTP</a>™) with 50 million cells in each vial that significantly reduces and/or eliminates the need for growing cells in the lab so that researchers in the field of tissue engineering and bioprinting can focus on their research rather than worry about optimizing their expansion protocols. At the development stage, this would significantly reduce the amount of time for researchers to conduct multiple experiments thus generating more data in a shorter time. To scale such a process for commercial production, it would be necessary to estimate the required manufacturing lot size as described in our <a href="http://roosterbio.blogspot.com/2018/09/building-effective-multi-year-process_12.html">previous blog</a> to build an effective multi-year process development program. In the case of the knee meniscus, there are over 700,000 patients requiring meniscus surgery in the US alone. To supply even 20% of this requirement, one would require 140,000 bioprinted knee meniscus which would be equivalent to 8.75 trillion cells annually.<br />
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<i>References:</i><br />
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1. Cohen et al., 2018. Tissue engineering the human auricle by auricular chondrocyte-mesenchymal stem cell co-implantation. PLoSOne13(10): e0202356. doi: 10.1371/journal.pone.0202356<br />
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2. Moller et al., 2017. In vivochondrogenesis in 3D bioprinted human cell-laden hydrogel constructs. Plast Reconstr Surg Glob Open. 5(2): e1227. Doi: 10.1097/GOX.000000000000127<br />
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</style>Sandy Applebeehttp://www.blogger.com/profile/01351576729478919332noreply@blogger.com0tag:blogger.com,1999:blog-6181073719515632715.post-30600296811272412572018-12-15T10:26:00.000-05:002019-06-26T14:53:54.243-04:00Building Effective Multi-Year Process Development Programs: Evolution of Technology Platform Decisions Based on Lot Size<i>Authored by Josephine Lembong, Ph.D., Scientist, Product & Process Development & Jon Rowley, Ph.D., Founder and Chief Product Officer, RoosterBio Inc.</i><br />
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Previously, we published a blog post on <a href="http://roosterbio.blogspot.com/2018/09/building-effective-multi-year-process_12.html">estimating hMSC lot sizes for clinical manufacturing</a>, with the goal of outlining a development program that could tailor accordingly. This exercise is crucial because the calculated target cell lot size dictates the final production platform needed for your therapeutic product. The next step would be to determine the appropriate manufacturing platform, for each unit operation, that will meet the calculated hMSC lot sizes throughout clinical development. Having a solid, multi-year plan will help your company succeed at navigating this complex maze that is the path to market success.<br />
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“It is the technologist’s and engineer’s jobs to drive the technology platform decision making process”</h2>
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The final decision of production platforms can be overwhelming; even though there is a certain goal in mind for the present time, you do want to keep it flexible and scalable for other potential applications in the future. There is also the goal of managing through the “Comparability Challenges” as these changes are implemented. Adding to the complexity is that as the <a href="https://en.wikipedia.org/wiki/Regenerative_medicine">RegenMed</a> industry grows, the technology providers of cell processing platforms across the various unit operations seems to be increasing in a fractal nature, with little standardization across devices. These technologies (e.g. bioreactors, continuous centrifuges, fill finish/controlled freezing, and other automation platforms) are significant investments to the company in the form of cost and time. It is the technologist’s and engineer’s jobs to drive the technology platform decision making process by derisking these technologies and establishing a multi-year development program, all while determining the costs associated with the program, and communicating these needs to the company’s business team so that they can raise the needed capital for these programs over time.<br />
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The goal of this post is to lay out the various scales of hMSC production and highlight the existing technology platforms for the different unit operations involved in the manufacturing process. This will help define the requirements that will guide the company’s multi-year process development program to meet projected future lot sizes.<br />
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<b><i><u>Each phase of a clinical trial is associated with a specific production scale, which dictates the production platform</u></i></b><br />
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At the end of the last <a href="https://roosterbio.blogspot.com/2018/09/building-effective-multi-year-process_12.html">post</a>, we arrived at the following estimated lot sizes based on a set of assumptions: 525 billion viable hMSCs per final commercial manufacturing lot, assuming a mid-range dose for a cardiac indication aimed to treat 100,000 patients per year, with a relatively safe, conservative assumptions regarding losses in cell viability and recovery during every step of the production process. Assuming a go-to-market lot size of 20% of the full commercial scale, we estimated that one could target a 100B cell lot size for Phase III, a 25B cell lot size for Phase II, and potentially a 10B cell lot size for a Phase I trial. These are simply guidelines that will change based on assumptions, but we recommend everyone go through this exercise (as outlined <a href="http://roosterbio.blogspot.com/2018/09/building-effective-multi-year-process_12.html">here</a>) for each therapeutic program.<br />
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Traditional 2D hMSC manufacturing processes were capable of harvest densities of 20,000-30,000 cells/cm2 [<a href="https://bioprocessintl.com/manufacturing/cell-therapies/meeting-lot-size-challenges-of-manufacturing-adherent-cells-for-therapy-328093/">1</a>]. Starting with a working cell bank of 40M cells, the Phase I lot size could be achieved using 60-80 2D CellStacks at this range of harvest density. High productivity hMSC Bioprocess systems such as <a href="https://www.roosterbio.com/collections/high-volume-hmscs">RoosterBio’s</a> drive cell density at harvest to ≥ 40,000 cells/cm2, while maintaining low cumulative PDL of ~16-18. The actual 2D harvest numbers can be > 60,000 cells/cm2 (this is mostly donor dependent), however downstream processing losses typically bring the numbers back down [<a href="https://pdfs.semanticscholar.org/3bb9/3813b696ff3f760476dc61a6ed228cdd85f3.pdf">2</a>], so we’d like to do all our calculations with a conservative density of 40,000 cells/cm2. This higher harvest density decreases the number of CellStacks to ~40 compared to traditional 2D planar manufacturing processes for this Phase I scale, reducing process complexity, labor and overall COGS significantly. Harvest processing time can be reduced by several hours and the facility footprint by hundreds of square feet, including many expensive reagents costs such as media, harvest reagent, etc. With these numbers, the processes can be developed and executed quickly in a robust 10-layer CellStack/CellFactory process, therefore getting a final hMSC product into Phase I for a proof to therapy with a low-risk process. Achieving a rapid and low cost Phase I feasibility is usually the best path for pharmaceutical development, and thus drives much of the 2D platform decisions for hMSC products. We do help our customers rapidly implement these processes via our <a href="https://www.roosterbio.com/pages/acceleration-services">Process Design and Acceleration Services</a>.<br />
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<b><i><u>Scalable hMSC clinical manufacturing needs movement away from 2D planar platforms</u></i></b><br />
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Each manufacturing phase leading up to the go-to-market lot size is associated with a new scale of production, thus requiring a scale-up development program. Process development programs are time-consuming and expensive, therefore justifying the necessity of a multi-year planning for efficient capital utilization within a company. Specific time-intensive planning aspects to proactively address include budgeting for at least 6 months of disposable engineering (having each single use bag and disposable at each unit operation with compatible connections), and then planning for at least 3-6 half scale runs and 3 full scale runs to understand and work out kinks in a full process prior to undertaking tech transfer. This can easily encompass up to 12-18 months of development studies for each scale-up program. <br />
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The scale of production at our proposed Phase II lot size of 25B is challenging to achieve using 2D planar platforms, even with high productivity systems. Assuming a typical harvest yield of 40,000 cells/cm2, 25B cells in CS10’s requires ~100 CellStacks. There are several challenges with this; it is very expensive, time consuming, labor intensive, and not environmentally friendly (think of the >800 sq ft footprint that the CellStacks occupy and how much plastic that is). The largest 2D planar platform, the automated 120-layer hyperstacks, can generate ~100B lot size, however there is no option of scaling-up from there. At scales larger than 15-20 Billion cells/lot, we recommend moving directly to 3D bioreactor platforms to maintain scalability for the future. We estimate that a 100B target lot size is relevant for ~95% of the indications requiring hMSC treatment based off of <a href="https://www.frontiersin.org/files/Articles/351606/fmed-05-00178-HTML/image_m/fmed-05-00178-t001.jpg">our analysis of cell dose</a> from the <a href="http://www.clinicaltrials.gov/">ClinicalTrials.gov</a> website [<a href="https://www.frontiersin.org/articles/10.3389/fmed.2018.00178/full">3</a>], with some exceptions including low dose indications such as ocular diseases [<a href="https://dx.doi.org/10.1002/bit.25008">4</a>]. Therefore, we recommend a jump to a bioreactor production platform for most development programs aiming to get to Phase II. <br />
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As the field moves into suspension bioreactors from 2D, there are new opportunities for productivity gains during production. Borrowing from best practices in the monoclonal antibody field, <a href="http://roosterbio.blogspot.com/2015/08/enabling-new-paradigm-in-hmsc.html#more">fed batch processes can lead to high media productivity</a>, and we have established this in <a href="https://roosterbio.blogspot.com/2015/08/enabling-new-paradigm-in-hmsc.html">hMSC production</a> as well. With a fed batch process, coupled with well QC’d, low PDL cell banks, it is routine to achieve a range of final cell yields of <a href="https://cdn.shopify.com/s/files/1/0515/3253/files/ISCT_Poster_XFR_Bioreactors_4.24.18_rev1___1_of_2.pdf?12483280270868979873">0.4M - 0.8M cells/ml</a> in a microcarrier suspension bioreactor process incorporating a <a href="https://www.roosterbio.com/products/roosterreplenish-msc-xf-su-023">bioreactor feed</a>, when performed in a low-shear <a href="https://www.pbsbiotech.com/products.html">Vertical-Wheel™ bioreactor</a>. A 3D fed-batch process increases media usage efficiency by <a href="https://cdn.shopify.com/s/files/1/0515/3253/files/ISCT_Poster_XFR_Bioreactors_4.24.18_rev1___1_of_2.pdf?3966769385012105074">>2 times compared to our 2D batch systems</a> (or by >15 times compared to a traditional hMSC 2D flask culture), thus simplifying the overall process, increasing cell productivity, and reducing media costs per cell produced. It is these compounded efficiencies <a href="https://www.frontiersin.org/files/Articles/351606/fmed-05-00178-HTML/image_m/fmed-05-00178-g009.jpg">that will drive the cell cost curve down</a> with further scale advancements.<br />
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For estimating bioreactor production volumes, we assume a conservative average bioreactor yield of 0.5M cells/ml (again, this is donor dependent, among other variables). Thus, a production vessel of 50L bioreactor is required for a 25B cell lot size; the 100B hMSC go-to-market lot size requires a 200L bioreactor, and the full-scale production requires a 1000L bioreactor. To achieve these lot sizes while maintaining the PDL of the final product (a critical quality attribute of every hMSC final product), the working cell bank must scale in size with the process, and will range from 100M to 2B (in closed, single use formats).<br />
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<span style="font-size: x-small;"><b>Table 1</b>. Summary of technology landscape for allogeneic hMSC manufacturing – by unit operation</span></div>
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<a href="https://2.bp.blogspot.com/-r9vex4i5bPQ/XBLagj7PcMI/AAAAAAAAAEg/-_VjqtLrLH49794Luvxzj0otSctFpcSnwCLcBGAs/s1600/blog%2B2018%2B12%2Btable%2B1.png" imageanchor="1" style="margin-left: 1em; margin-right: 1em;"><img border="0" data-original-height="532" data-original-width="1600" height="212" src="https://2.bp.blogspot.com/-r9vex4i5bPQ/XBLagj7PcMI/AAAAAAAAAEg/-_VjqtLrLH49794Luvxzj0otSctFpcSnwCLcBGAs/s640/blog%2B2018%2B12%2Btable%2B1.png" width="640" /></a></div>
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<i><b><u>Proactively addressing downstream processing bottlenecks</u></b></i><br />
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The subsequent post-harvest processing steps involve unit operations which rely on the scale of the expansion platform [<a href="https://pdfs.semanticscholar.org/3bb9/3813b696ff3f760476dc61a6ed228cdd85f3.pdf">2</a>], therefore it is critical to proactively address downstream process unit operations as you choose your expansion technologies. You don’t want to manufacture your products at a scale only to waste a large fraction of it due to incapability to process. Cell harvesting from the bioreactor is immediately followed by a cell-microcarrier separation process. While there are no standard accepted criteria for the amount of microcarriers that can be in the final product, near complete removal of microcarriers from the cell therapy product is critical in ensuring safety for the patient. For the microcarrier removal unit operation, screen-based depth filtration technology has been implemented, trapping microcarriers within 60-100 µm meshes while letting cells pass through. This leaves the cells in the quenched harvest reagent for further clarification and concentration. Alternatively, the development and use of <a href="https://www.corning.com/worldwide/en/products/life-sciences/applications/bioprocess-applications/cell-therapy/dissolvable-microcarriers.html">dissolvable microcarriers</a> in large scale hMSC bioreactor culture could potentially eliminate the need for a separation unit, creating a more streamlined, simplified hMSC manufacturing process.<br />
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<a href="https://3.bp.blogspot.com/-7BONgTxQEBI/XBLJ8b6Kx7I/AAAAAAAAAEI/zj9LqwX15ikxpcY45hpgLwY-AOJK5lOfQCLcBGAs/s1600/blog%2B2018%2B12%2Bfigure%2B3.png" imageanchor="1" style="margin-left: 1em; margin-right: 1em;"><img border="0" data-original-height="499" data-original-width="1600" height="199" src="https://3.bp.blogspot.com/-7BONgTxQEBI/XBLJ8b6Kx7I/AAAAAAAAAEI/zj9LqwX15ikxpcY45hpgLwY-AOJK5lOfQCLcBGAs/s640/blog%2B2018%2B12%2Bfigure%2B3.png" width="640" /></a></div>
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The microcarrier-free cell suspension coming out of the separation step goes into a volume reduction / wash operation. Large scale production of hMSCs (starting at Phase II) demands scalable technologies for this step, i.e. tangential flow filtration (TFF) or continuous counterflow centrifugation system [<a href="https://pdfs.semanticscholar.org/3bb9/3813b696ff3f760476dc61a6ed228cdd85f3.pdf">2</a>] such as <a href="https://www.sartorius.com/en/products/process-filtration-purification/process-filtration-harvesting/ksep-systems">kSep</a> (Sartorius) and <a href="https://www.scinogy.com/rotea/">Rotea</a> (Scinogy). Newer technologies such as acoustic wave separation technology (e.g. <a href="https://shop.pall.com/us/en/biotech/continuous-processing/clarification/cadence-acoustic-separator-zidimmfdb9d">Pall’s Cadence™ Acoustic Separator</a>, <a href="https://www.fdsonics.com/">FloDesign Sonics</a>) are also being developed and optimized for hMSC manufacturing process. With 100B lot size, the kSep is the only platform today that can accommodate 200L volume within a reasonable manufacturing timescale (to give a rough idea, our experience of processing of 60L of 0.4M cells/ml in the kSep400 with conservative process parameters takes about 1.5 hours in one single cycle, while a typical open centrifugation system for this volume would have taken over 3 hours in 30 consecutive cycles). At 500B cells, the kSep6000S is the only platform that could potentially process the volume associated with this full commercial scale lot size. Other volume reduction technologies such as Rotea or FloDesign Sonics’ Acoustic Wave Technology can be implemented in productions associated with earlier phases or indications associated with smaller lot sizes due to their smaller processing capabilities (10-50B cells), however keep in mind that these technologies might scale in the future for hMSC manufacturing. Studies can also be done to test these systems for a combined separation / concentration step during the manufacturing steps, but this will require extensive process development and validation for use with cell/microcarrier suspensions.<br />
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The envisioned format of the final cell product is a pharma-style vial with ~75M cells/vial at 10 ml fill. To fill 10B cells, a semi-automated vialing system can be used such as the <a href="https://www.aseptictech.com/products/m1-filling-station">M1 Filling Station</a> (Aseptic Technologies). For the full scale production at 500+ vials/hr, a fully automated vialing system such as the <a href="https://www.aseptictech.com/products/robot-lines">Crystal® L1 Robot Line</a> can be employed. <br />
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Cryopreservation, followed by storage, is the last step of the manufacturing process. Since fast, controlled rate freezing process is critical in maintaining cell health, large-scale cryopreservation platforms such as the <a href="https://www.thermofisher.com/order/catalog/product/7450">CryoMed™ Controlled Rate Freezers</a> (CRF) which allows a custom user-defined recipe is commonly used. Each process development group will work to optimize a controlled rate freezer recipe that works for their final products, with post-thaw viability and expansion being the performance metric.<br />
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After all processes, it is important to verify the final cell product quality. The safety of the cells needs to remain intact and <a href="http://www.roosterbio.com/pages/BPI-poster-2018">critical quality attributes of the cells need to be maintained</a>. RoosterBio performs a general analytic panel to ensure safety and critical quality attributes of hMSCs (including cell performance, identity, and functionality testing), but each final product will have its unique testing requirements and must be performed within the context of each development program. However, our goal is to identify and derisk the comparability challenges for our customers prior to them scaling, simplifying and accelerating the process and product development for each intended program.<br />
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In conclusion, we have highlighted various existing technology platforms involved in an hMSC manufacturing process. You can think of this exercise as a decision-making tool to help streamline an hMSC clinical manufacturing program. As this informs the development team on the unit operation choices associated with each phase, it also helps the team define their requirements and invest in lot-size-appropriate technologies. This leads to a RegenMed company’s multi-year planning of its process development program to meet the peak commercial demands for most indications requiring hMSC treatment.<br />
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If you have an interest in discussing scalable production platforms, evaluating our complete solutions that radically simplify hMSC process development, or are interested in our <a href="https://www.roosterbio.com/pages/acceleration-services">Process Design & Acceleration Services</a>, please reach out to us at info@roosterbio.com, or simply comment on this blog post!<br />
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<i><u>References:</u></i><br />
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<li>Rowley et al., 2012. Meeting Lot-Size Challenges of Manufacturing Adherent Cells for Therapy. BioProcess International 10(3): 16-22</li>
<li>Pattasseril et al., 2013. Downstream Technology
Landscape for Large-Scale Therapeutic Cell Processing. <i>BioProcess International</i> 11(3): 38-47</li>
<li>Olsen et al., 2018. Peak MSC—Are We There Yet?
Front Med 5:178. doi: 10.3389/fmed.2018.00178</li>
<li>Simaria et al., 2013. Allogeneic cell therapy
bioprocess economics and optimization: Single‐use cell expansion technologies. <i>Biotechnol Bioeng</i> 111(1): 69-83. doi:
10.1002/bit.25008</li>
</ol>
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Anonymousnoreply@blogger.com0tag:blogger.com,1999:blog-6181073719515632715.post-5838441178311080352018-11-17T09:47:00.001-05:002019-06-11T16:37:45.097-04:00RoosterBio Products Continue to Expand Presence in Major Mesenchymal Stem/Stromal Cell (MSC)-related Regenerative Medicine Peer Reviewed Publications<i>Authored by Joseph Candiello, Ph.D., Technical Application Specialist</i><br />
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<tr><td class="tr-caption" style="text-align: center;">Figure 1. Current
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Human mesenchymal stem/stromal cells (hMSCs) are a critical starting material in a growing variety of established and emerging applications spanning the Regenerative Medicine space (Fig1). Their unique balance of functional bioactive attributes, expansion potential, and established safety profile have resulted in a steady increase in both <a href="https://bioinformant.com/comparison-of-stem-cell-research-frequency-by-stem-cell-type/">scientific publications</a> and a parallel increased presence in <a href="https://www.frontiersin.org/articles/10.3389/fmed.2018.00178/full">clinical application</a>s over the past 10+ years. Specific to scientific publications, from 2011 to 2015 there were <a href="https://bioinformant.com/two-thirds-of-all-mesenchymal-stem-cell-msc-publications-were-released-in-the-past-5-years/">over 17,000 MSC related publications</a>, with over 50% pertaining to human MSC use. This trend has continued with over 9,600 MSC related publications in 2016-2017 and with over 5,000 articles using human hMSCs. <br />
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We are excited that over the past 4 years RoosterBio’s core technology, hMSC cell banks and associated Bioprocess Media Systems have been a used in an increasing number of these peer reviewed scientific publications or what we affectionally refer to as our customer’s <a href="https://www.roosterbio.com/pages/the-brainy-stuff">“brainy stuff”</a>. The initial articles using RoosterBio technology were published in <a href="https://pubs.acs.org/doi/abs/10.1021/acs.biomac.5b00940">Biomacromolecules</a> and <a href="https://www.ncbi.nlm.nih.gov/pubmed/26139547">Cytotherapy</a> in the 2nd half of 2015; less than 2 years from when our first products left RoosterBio’s doors. During 2016 and 2017, RoosterBio products have appeared in leading academic journals such as <a href="http://www.pnas.org/content/113/12/3179">PNAS</a> <a href="https://www.ncbi.nlm.nih.gov/pubmed/28283659">(twice)</a>, <a href="https://www.ncbi.nlm.nih.gov/pubmed/27019026">Biomaterials</a>, <a href="https://www.ncbi.nlm.nih.gov/pubmed/28170190">Stem Cells and Translational Medicine</a>, <a href="https://www.ncbi.nlm.nih.gov/pubmed/28853608">American Journal of Respiratory and Critical Care Medicine</a>, and <a href="https://www.ncbi.nlm.nih.gov/pubmed/28853608">Nucleic Acids Research</a>.</div>
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In addition to a presence in these high-profile publications, RoosterBio's hMSC manufacturing-focused products, have had a steady yearly increase in overall total peer reviewed articles in print (Fig. 2). In 2018 to date a total of 35 articles have included RBI products, surpassing 2017’s total and doubling 2016’s output. In addition to total publications, we have tracked articles using a few key metrics – Journal’s Impact Factor (IF), an index of citations per publication, and studies which contain an in vivo, or animal model component (Fig 2). With respect to IF, articles using RBI product have had an increase in average IF from 4.4 in 2016 to 5.2 in 2018 and total number of publications with an IF >4 from 5 articles in 2015 to 17 to date in 2018, which is almost half of the total articles this year. Original research articles with an in vivo component have had similar increases, from 2 articles (12%) in 2015 to 10 articles (28%) so far in 2018. <br />
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The breadth of topics included in these publications is as diverse as <a href="https://www.frontiersin.org/articles/10.3389/fmed.2018.00178/full">current applications across the hMSC Regenerative Medicine space</a> – and is called out in our <a href="https://cdn.shopify.com/s/files/1/0515/3253/files/RoosterReference_Database_NOV2018_Website_NoData.xlsx?10927442456194162988">databaseof current RoosterBio product publications</a>. As frequency of publications increases, we will highlight some of these – starting with some recent exciting work being reported across the industry:</div>
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<a href="https://www.blogger.com/blogger.g?blogID=6181073719515632715" imageanchor="1" style="clear: right; float: right; margin-bottom: 1em; margin-left: 1em;"></a><b>As RoosterBio continues to provide hMSC Bioprocess Systems to support Regenerative Medicine research and product development efforts; the central focus is on how we can radically simplify our customers workflow. To this end, across the life sciences industry, timeframes from project initiation to first publication continue to increase; with a current <a href="http://www.pnas.org/content/112/44/13439">average of 3 to 4 years</a>. At the same time, it is critical for researchers and product developers to shorten both publication and product development timeframes. </b><a href="https://www.roosterbio.com/pages/products">RoosterBio high volume hMSCs and bioprocess media systems</a> are uniquely designed to accelerate these timeframes by providing well characterized, consistent raw materials designed for simplicity, reduced cost, and shortened time to generate the necessary cellular components for research, development, or clinical translation.<br />
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Highlighted Publications:<br />
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<a href="https://www.ncbi.nlm.nih.gov/pubmed/30384124">Defining Hydrogel Properties to Instruct Lineage- and Cell-Specific Mesenchymal Differentiation</a>. Hung BP, Harvestine JN, Saiz AM, Gonzalez-Fernandez T, Sahar DE, Weiss ML, Leach JK, <b>Biomaterials</b>, 2019.<br /><br /><a href="https://www.ncbi.nlm.nih.gov/pubmed/29352647">IFN-γ and TNF-α Pre-licensing Protects Mesenchymal Stromal Cells from the Pro-inflammatory Effects of Palmitate</a>. Boland L, Burand AJ, Brown AJ, Boyt D, Lira VA, Ankrum JA. <b>Mol Ther.</b> 2018.<br /><br /><a href="https://www.ncbi.nlm.nih.gov/pubmed/29693099">Deciphering the role of substrate stiffness in enhancing the internalization efficiency of plasmid DNA in stem cells using lipid-based nanocarriers</a>. Modaresi S, Pacelli S, Whitlow J, Paul A. <b>Nanoscale. </b>2018.<br /><br /><a href="https://www.ncbi.nlm.nih.gov/pubmed/30182058">Acoustophoretic printing</a>. Foresti D, Kroll KT, Amissah R, Sillani F, Homan KA, Poulikakos D, Lewis JA. <b>Sci Adv.</b> 2018.<br /><br /><a href="https://www.ncbi.nlm.nih.gov/pubmed/30248646">3D printed biofunctionalized scaffolds for microfracture repair of cartilage defects.</a> Guo T, Noshin M, Baker HB, Taskoy E, Meredith SJ, Tang Q, Ringel JP, Lerman MJ, Chen Y, Packer JD, Fisher JP. <b>Biomaterials.</b> 2018.<br /><br /><a href="https://www.ncbi.nlm.nih.gov/pubmed/29251710">Mesenchymal stem cell-derived extracellular vesicles attenuate pulmonary vascular permeability and lung injury induced by hemorrhagic shock and trauma.</a> Potter DR, Miyazawa BY, Gibb SL, Deng X, Togaratti PP, Croze RH, Srivastava AK, Trivedi A, Matthay M, Holcomb JB, Schreiber MA, Pati S. J <b>Trauma Acute Care Surg</b>. 2018.<div class="MsoNormal" style="margin-left: .5in; text-align: justify; text-indent: -.5in;">
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If you are interested in having a conversation about how RoosterBio can shorten your gaps between experiments and accelerate your time to publication, contact us at info@roosterbio.com.</div>
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<br />Joe Candiellohttp://www.blogger.com/profile/06264220321055111520noreply@blogger.com0tag:blogger.com,1999:blog-6181073719515632715.post-12892200867649417882018-09-12T09:54:00.001-04:002019-06-11T16:46:22.657-04:00Building Effective Multi-Year Process Development Programs: Estimating hMSC Lot Size Ranges for Clinical Manufacturing through Commercial Demand – it is all about the assumptions<i>Authored by </i><i>Josephine Lembong, Ph.D., Scientist, Product & </i><i>Process </i><i>Development & </i><i>Jon A. Rowley, Ph.D., Founder and Chief Product Officer, RoosterBio Inc.</i><br />
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<span style="font-family: inherit;"><a href="https://www.roosterbio.com/pages/jon-rowley" target="_blank">Jon Rowley</a>, <a href="https://www.roosterbio.com/" target="_blank">RoosterBio</a>’s Founder and Chief Product Officer, gave a talk at <a href="http://isct2018.com/" target="_blank">ISCT 2018</a> in May and had a set of slides about estimating lot size that had a lot of people scrambling to take notes. We thought we would share the content here at the RoosterBio blog with more discussion and make the content broadly available and open for discussion. </span><br />
<span style="font-family: inherit;"><br /></span>
<span style="font-family: inherit;">The concept is consistent with all good strategic planning; </span><br />
<ol>
<li><span style="font-family: inherit;">Understand what the future looks like and work backwards,</span></li>
<li><span style="font-family: inherit;">Create a model using reasonable to conservative assumptions along the way to estimate a range of needs,</span></li>
<li><span style="font-family: inherit;">Use this model to lay out what the next several years will look like assuming successful achievement of intermediate milestones, and create multi-year programs that are right-sized, with the right technology platform, for the stage of product / clinical development of your company.</span></li>
</ol>
<span style="font-family: inherit;">This is the type of consultation that we provide in our </span><a href="https://www.roosterbio.com/pages/acceleration-services" style="font-family: inherit;" target="_blank">Process Design and Acceleration</a><span style="font-family: inherit;"> business unit at RoosterBio, which we offer to our customers that are incorporating </span><a href="https://www.roosterbio.com/collections/high-volume-hmscs" style="font-family: inherit;" target="_blank">RoosterBio hMSC cell banks and bioprocess media systems</a><span style="font-family: inherit;"> into their product and process development efforts. However, we love to share our knowledge with the broader RegenMed Industry and are offering it up here.</span><br />
<span style="font-family: inherit;"><br /></span>
<span style="font-family: inherit;">Notes to readers: this exercise has the base assumption that this is an Allogenic, Universal Donor manufacturing process [<a href="https://doi.org/10.1002/bit.25008" target="_blank">1</a>] and is not applicable to Autologous or patient-specific products. The core logic would still hold when applied to autologous, but at a much smaller scale.</span><br />
<span style="font-family: inherit;"><br /></span>
<span style="font-family: inherit;"><b><i><u>Lot Size Targets Dictate the Production Platform </u></i></b></span><br />
<span style="font-family: inherit;"><br /></span>
<span style="font-family: inherit;">The goal of any process development program is to create a right-sized manufacturing process for your immediate business goals, but with future scalability in mind. Understanding the future lot size requirements will help strategically align manufacturing requirements today with commercial scalability while laying a platform foundation to minimize comparability risks.</span><br />
<span style="font-family: inherit;"><br /></span>
<span style="font-family: inherit;"><b><i><u>First Start with the End in Mind – Engage with Marketing</u></i></b></span><br />
<span style="font-family: inherit;"><br /></span>
<span style="font-family: inherit;">Get with the business team within your company (usually Strategic Marketing and/or Commercial Operations) and request a numbers-driven discussion on which of the multiple indications that a process development program should be built for. There are usually several therapeutic indications of interest, but it is important to realize that a manufacturing process for an ocular indication that has a dose of 1 million cells/dose is very different than a Crohn's disease indication that has a dose size of greater than 1 billion cells/dose [<a href="https://doi.org/10.3389/fmed.2018.00178" target="_blank">2</a>]. Each therapeutic indication from a company must be treated as a distinct product and target, with a distinct process. Just because the same cell type is used in both therapeutic products does not mean they share a manufacturing process. This is critical to bring clarity to each program.</span><br />
<span style="font-family: inherit;"><br /></span>
<span style="font-family: inherit; font-size: large;"><i>“A common pitfall that many cell therapy companies fall into is that each therapeutic indication is not addressed as a unique product with a unique manufacturing process”</i></span><br />
<span style="font-family: inherit;"><br /></span>
<span style="font-family: inherit;">Once the team is aligned on the target indication (or set of prioritized target indications), then you need to understand what a reasonable peak commercial market demand would be – this is likely to be found in the business model projections that are built for investors and is a good place to start. If there are 500,000 patients a year in your target indication and the business is aiming to capture 20% of them (or treat 100,000 patients/year) at peak commercial demand, then you need a process that will scale with that over time, assuming both clinical and market success. It is a good idea to establish a range. We often like to perform this exercise with a low, middle and high assumptions (such as 30,000 patients as low, 100,000 as the target, and 250,000 patients treated/year if wildly successful). For this article, we will simply focus on the '100,000 patients treated assumption' for our calculations.</span><br />
<span style="font-family: inherit;"><br /></span>
<span style="font-family: inherit;"><b><u><i>Dosing Assumptions: this is one of the hardest parts</i></u></b></span><br />
<span style="font-family: inherit;"><br /></span>
<span style="font-family: inherit;">Estimating dosing is sometimes the most difficult part. Does the patient need a single dose or multiple doses, and how many cells per dose? In many cases a dose escalation trial has not yet been performed, so some educated guesstimates are required. Taking a low/mid/high range of estimates also works here, and there is sufficient published work related to hMSCs in different indications that you can get close enough for these purposes [</span><a href="https://doi.org/10.3389/fmed.2018.00178" target="_blank">2</a><span style="font-family: inherit;">,</span><a href="https://doi.org/10.3727%2F096368915X689622" style="font-family: inherit;" target="_blank">3</a><span style="font-family: inherit;">]. For this example, we will assume a cardiac indication. There is a good amount of published work on hMSCs for cardiac indication and we can assume a conservative low dose of 25 million cells, a mid-range dose of 50 million, and a high dose of 100 million [</span><a href="https://doi.org/10.1002/sctm.16-0484" style="font-family: inherit;" target="_blank">4</a>,<a href="https://doi.org/10.5966/sctm.2015-0101" target="_blank">5</a><span style="font-family: inherit;">]. In this blog post we will perform all calculations with an assumption of 50 million cells per dose.</span><br />
<span style="font-family: inherit;"><br /></span>
<span style="font-family: inherit;"><b><u><i>Calculating Yearly Product Needs at Peak Commercial Demand</i></u></b></span><br />
<ul>
<li>At peak demand, we aim to produce 100,000 doses per year, with a size of 50 million cells per dose.</li>
<li>100,000 doses per year ≈ 2,000 doses per week</li>
<ul>
<li>Weekly production is not recommended – it is simply unsustainable, so let’s assume one (1) lot production every two (2) weeks, or 25 lots total in a year (which is still a lot for any manufacturing site)</li>
<li>Not all lots will be successful – assume a 10% scrap rate, thus 2-3 lots are lost per year, giving us 22-23 lots per year</li>
<li>100,000 doses per year / ~23 lots per year = 4,450 doses/lot </li>
<li>Assume 10% of lot goes to testing (~450 dose), so a final target lot size of <b>4,900 doses/lot will be needed to be manufactured at peak commercial demand</b></li>
</ul>
</ul>
<span style="font-family: inherit;">It is important to remember that it typically takes multiple years for a successful therapeutic to reach peak demand, so it is not necessary to go to market with a manufacturing process capable of meeting peak demand – especially for an early stage field like <a href="https://en.wikipedia.org/wiki/Regenerative_medicine" target="_blank">Regenerative Medicine</a>. We will create estimates for peak demand, but then focus on building a reasonable “go-to-market” lot size and manufacturing process.</span><br />
<span style="font-family: inherit;"><br /></span>
<span style="font-family: inherit;"><b><u><i>Calculating Total Cells to Manufacture that are Required to Fill into the Final Container</i></u></b></span><br />
<ul>
<li>We will assume 1 dose goes into one container.</li>
<li>We assumed a mid-range dose size of 50 million (i.e. 50 million viable cells per vial, post-thaw).</li>
<li><a href="https://chemometec.com/applications/cell-counting-cell-therapeutic-applications/" target="_blank">Cell Count</a> is a strict QC parameter with a straight forward specification. It is important to point out that often there is a solid safety factor, or overfill, applied to this metric. You never want to fail a lot on cell count after you spent hundreds of thousands of dollars creating the units – so the risk averse will overfill vials until sampling and cell enumeration is an accurate, consistent, and precise method.</li>
<ul>
<li>Assume 15% (worst case) viability drop and 20% recovery loss during cryopreservation (these assumptions should be data driven from your process, and constantly trended and updated during manufacturing).</li>
<li>Based on the above assumptions, to consistently achieve 50 million viable cells after thaw, it is required to target filling at <b>75 million viable cells</b> into the final container:</li>
<ul>
<li>75 million viable cells – 15 million (20% total cell loss on recovery) = 60 million viable cells </li>
<li>60 million – 9 million (15% viability drop) = 51 million viable cell target specification</li>
</ul>
</ul>
<li>NOTE: some programs will only overfill by 10-20%, but we find that with this range it is very difficult to routinely obtain successful cell counts in a manufacturing/QC environment, so we recommend 30-50% overfill for calculations.</li>
</ul>
<b style="font-family: inherit;"><u><i><br /></i></u></b>
<b style="font-family: inherit;"><u><i>Calculating Lot Size Requirements at Peak Commercial Demand</i></u></b><br />
<span style="font-family: inherit;"><br /></span>
<span style="font-family: inherit;">One unfortunate fact of cell manufacturing is that you need to manufacture more cells than you need to fill, since there will be losses associated with the post-harvest processing steps. Many biological manufacturing processes have losses in the 40%-60% range in this “downstream” processing. For our process, we go through the following math:</span><br />
<ul>
<li>75 million cells/dose × 4900 doses per manufacturing lot = 368 billion viable cells needed to target 4900 doses from every manufacturing lot.</li>
<li>We assume 30% post-harvest cell loss during processing for this exercise – thus at least <b>525 billion cells need to be manufactured</b> in order to have 368 billion cells during the fill unit operation.</li>
<ul>
<li>525 billion viable cells – 157 billion (30% post-harvest cell loss) = 368 billion viable cells </li>
</ul>
<li>30% cell loss does seem significant, however every process development group will have a team working to optimize their process to minimize this number. It is likely that recoveries from earlier processes will be much greater than 30%, and with optimization at large scale it is possible to improve. For modeling, it is always worth using reasonable to conservative estimates.</li>
</ul>
In conclusion, we arrived at the following numbers: <br />
<ul>
<li>At peak commercial demand, 23 successful lots of 525 billion cells per lot need to be manufactured in order to meet the demand, assuming a mid-range dose for a cardiac indication aimed to treat 100,000 patients per year. </li>
</ul>
This calculation does not take into account further downstream losses associated with stability, multiple distribution sites, safety stocks – which will just make the numbers worse. For this exercise, we will stop here. The 525 billion viable cells per lot is the number we were looking for, and it is this number that will drive the production platform decisions for streamlined hMSC clinical manufacturing.<br />
<br /> For an early-stage RegenMed company, a go-to-market lot size of ~20% of the “peak commercial scale” is a reasonable target to plan for, so a ~100 billion cells/lot would be a good target number to develop and run your Phase III trial at.<span style="font-family: inherit;"><br /></span>
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<u><i><b>We’d like to summarize this post with a few number recommendations:</b></i></u><div class="MsoNormal">
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<br />Phase III/Go-to-Market: ~100B cells<br /><br />Phase II (100-200 patients over 3-4 lots): 15-25B cells – this is an intermediate step up from Phase I and within a log of the Phase III/go-to-market process.<br /><br />Phase I (small scale): 5-10B cells <br /><br />The next stage is to lay out the technology platforms for the various manufacturing unit operations that are capable of processing these cell numbers. This will be the topic of our next blog post.<br /><br /><i>References: </i><br /><ol>
<li>Simaria et al., 2013. Allogeneic cell therapy bioprocess economics and optimization: Single‐use cell expansion technologies. Biotechnol Bioeng 111(1): 69-83. doi: 10.1002/bit.25008 </li>
<li>Olsen et al., 2018. Peak MSC—Are We There Yet? Front Med 5:178. doi: 10.3389/fmed.2018.00178 </li>
<li>Squillaro et al., 2016. Clinical Trials With Mesenchymal Stem Cells: An Update. Cell Transplant 25(5):829-848. doi: 10.3727/096368915X689622 </li>
<li>Majka et al., 2017. Concise Review: Mesenchymal Stem Cells in Cardiovascular Regeneration: Emerging Research Directions and Clinical Applications. Stem Cells Transl Med 6(10):1859-1867. doi: 10.1002/sctm.16-0484 </li>
<li>Golpanian et al., 2016. Concise Review: Review and Perspective of Cell Dosage and Routes of Administration From Preclinical and Clinical Studies of Stem Cell Therapy for Heart Disease. Stem Cells Transl Med 5(2):186-191. doi: 10.5966/sctm.2015-0101</li>
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Anonymousnoreply@blogger.com0tag:blogger.com,1999:blog-6181073719515632715.post-70884651140791075302018-05-16T14:16:00.001-04:002019-05-10T16:00:48.004-04:00ISCT 2018: MSC Biomanufacturing, Bioprocessing, Scale-Up, Analytics and Exosome Production Take Center Stage<span style="font-family: "arial" , "helvetica" , sans-serif; font-size: x-small;"><br /></span>
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<span style="font-family: "arial" , "helvetica" , sans-serif;"><i>Authored by Katrina Adlerz, Ph.D. Scientist, Analytical Development, RoosterBio Inc.</i></span><br />
<span style="font-family: "arial" , "helvetica" , sans-serif;"><br /></span>
<span style="font-family: "arial" , "helvetica" , sans-serif;">RoosterBio attended the <a href="http://www.celltherapysociety.org/default.asp?">International Society for Cell Therapy</a> annual meeting, held
May 2-5, 2018 in Montreal, which brought together leaders in the field including
academic researchers, industry scientists, regulators, and clinicians. The
society and conference focus on three key areas of translation: Academia,
Regulatory, and Commercialization.<o:p></o:p></span></div>
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<a href="https://4.bp.blogspot.com/-uboqe7rXXo8/WvxM_xIM1hI/AAAAAAAAFN4/E91YxGSucvAwmJvXbPN-YfE1vrFr5VhCgCLcBGAs/s1600/RoosterBio%2BBooth.jpg" imageanchor="1" style="clear: left; float: left; margin-bottom: 1em; margin-right: 1em;"><span style="font-family: "arial" , "helvetica" , sans-serif;"></span></a><span style="font-family: "arial" , "helvetica" , sans-serif;">Six Roosters attended to hear the latest
research and translational innovations at the scientific talks, present three
different posters, and man the booth in the exhibit hall. <a href="https://www.roosterbio.com/pages/about-us" target="_blank">Jon Rowley, founder and CTO of RoosterBio,</a> also gave two talks discussing innovations to accelerate
MSC Biomanufacturing, Bioprocessing and Scale-Up. He shared insights from RoosterBio’s path to <a href="https://www.roosterbio.com/pages/clinicontrol-products" target="_blank">GMP-manufactured cells and media</a> as well as strategies for overcoming obstacles in his talk “<b>Technologies for Radically Reducing
Development Timelines of hMSC-based Therapeutic Products</b>” during the
<a href="http://isct2018.com/program/" target="_blank">Strategies </a><a href="http://isct2018.com/program/" target="_blank">for </a><a href="http://isct2018.com/program/" target="_blank">Commercialization Session</a>. (New to MSCs? Read more about MSCs and biomanufacturing <a href="http://roosterbio.blogspot.com/2014/02/what-are-mscs.html" target="_blank">here</a>, <a href="http://roosterbio.blogspot.com/2014/02/current-bottlenecks-in-msc-research.html" target="_blank">here</a>, <a href="http://roosterbio.blogspot.com/2014/03/mesenchymal-stem-cells-workhorse-of.html" target="_blank">here</a> and <a href="http://roosterbio.blogspot.com/2017/06/opportunities-for-biomanufacturing.html#more" target="_blank">here</a>.) For a copy of Jon's talk, <a href="mailto:priya@roosterbio.com" target="_blank">email us</a>.</span></div>
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<span style="font-family: "arial" , "helvetica" , sans-serif;">RoosterBio Analytical, Process & Product Development efforts were well-represented with three
posters.</span></div>
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<b style="mso-bidi-font-weight: normal;">“A Xeno-Free Fed-Batch Microcarrier
Suspension Bioreactor System for the Scalable and Economic Expansion of
hBM-MSCs”</b> showed that MSC critical quality attributes were maintained in
0.1L and 3L bioreactor culture. <a href="https://cdn.shopify.com/s/files/1/0515/3253/files/ISCT_Poster_XFR_Bioreactors_4.24.18_rev1___1_of_2.pdf?12483280270868979873" target="_blank">Poster</a>.<o:p></o:p></span></div>
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<v:shape alt="" id="_x0000_s1027" style="height: 49.15pt; left: 0; margin-left: 389.25pt; margin-top: 2.7pt; mso-height-percent: 0; mso-height-percent: 0; mso-position-horizontal-relative: text; mso-position-vertical-relative: text; mso-width-percent: 0; mso-width-percent: 0; mso-wrap-edited: f; position: absolute; text-align: left; width: 65.5pt; z-index: 251663360;" type="#_x0000_t75" wrapcoords="-208 0 -208 21046 21600 21046 21600 0 -208 0">
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</xml><![endif]--><span style="font-family: "arial" , "helvetica" , sans-serif;">- “<b style="mso-bidi-font-weight: normal;">Scalable Xeno-Free Manufacturing of
Extracellular Vesicles Derived from Human Mesenchymal/Stromal Stem Cells</b>”
explained a process for generating a high yield of EVs/Exosomes from MSCs in a shortened
time frame using RoosterNourish<sup>TM</sup>-MSC-XF Media. <a href="https://cdn.shopify.com/s/files/1/0515/3253/files/ISCT_Poster_EV_Apr2018___2_of_2.pdf?12483280270868979873" target="_blank">Poster</a>.<o:p></o:p></span></div>
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</xml><![endif]--><span style="font-family: "arial" , "helvetica" , sans-serif;">- “<b style="mso-bidi-font-weight: normal;">Development & Technology Transfer of a cGMP
Potency Assay: Testing of an Ancillary Material for Stem Cell Manufacturing</b>”
outlined the steps in assay development, assay qualification/validation, and
tech transfer for a custom potency assay based on cell expansion. <a href="https://cdn.shopify.com/s/files/1/0515/3253/files/20180430_ISCT_2018_Poster___AD_Poster_3_of_3.pdf?12483280270868979873" target="_blank">Poster</a>.</span></div>
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<span style="font-family: "arial" , "helvetica" , sans-serif;">- In addition, the RoosterBio , BioLife Solutions and Brooks Life Science teams collaboratively presented a poster on MSC cryopreservation: <b>"</b><span style="font-weight: bold;">The Effect of Cryomedia Selection and Transient Warming Events on Post-Cryopreservation </span><b>Human MSC Function". </b><a href="https://cdn.shopify.com/s/files/1/0515/3253/files/BioLife-Brooks-RoosterBio_ISCT_Poster_v3_5-1-18.pdf?12483280270868979873" target="_blank">Poster</a>.<b> </b></span></div>
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<span style="font-family: "arial" , "helvetica" , sans-serif;">The posters gave presenting scientists the
opportunity to talk with those doing similar work in process or assay
development. It allowed us to learn from and share our expertise with the
community.</span></div>
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<span style="font-family: "arial" , "helvetica" , sans-serif;">The conference kicked
off with a <a href="http://isct2018.com/mscworkshop/" target="_blank"><b>RoosterBio-sponsored</b> </a><b style="mso-bidi-font-weight: normal;"><a href="http://isct2018.com/mscworkshop/" target="_blank">workshop</a>: Improving Mesenchymal Stem Cell Potency and Survival</b>. Steven Bauer, Branch Chief of the Cellular and Tissue Therapy Branch of the <a href="https://www.fda.gov/AboutFDA/CentersOffices/OfficeofMedicalProductsandTobacco/CBER/" target="_blank">US FDA</a>, and Head of the FDA's <a href="https://www.fda.gov/forconsumers/consumerupdates/ucm393030.htm" target="_blank">MSC Consortium</a>, discussed his innovative work developing predictive assays for MSC potency by
analyzing cell morphology in his talk “High Throughput Approaches to Assess MSC
Function”. You can find his blog <a href="https://blogs.fda.gov/fdavoice/index.php/tag/fdas-msc-consortium/" target="_blank">here</a>. </span><span style="font-family: "arial" , "helvetica" , sans-serif;">There were also a number of talks discussing clinical trials
results. </span><b style="font-family: Arial, Helvetica, sans-serif;">A common theme of the session
was the need to develop analytical methods and assays that can predict MSC-based treatment efficacy in
patients.</b><br />
<a name='more'></a><b style="font-family: Arial, Helvetica, sans-serif;"> </b><span style="font-family: "arial" , "helvetica" , sans-serif;">In addition to cell-based assays, predicting which patient
populations will respond to treatment would be a big step forward and likely
result in increased success with regards to patient outcomes. Eleuterio Lombardo
of </span><span style="font-family: "arial" , "helvetica" , sans-serif;"> </span><a href="http://tigenix.com/" style="font-family: Arial, Helvetica, sans-serif;" target="_blank">TiGenix</a><span style="font-family: "arial" , "helvetica" , sans-serif;"> discussed the development of such
a test during a clinical trial, where they found a group of 15 </span><a href="https://www.sciencedirect.com/topics/neuroscience/biomarkers" style="font-family: Arial, Helvetica, sans-serif;" target="_blank">biomarkers</a><span style="font-family: "arial" , "helvetica" , sans-serif;"> that
they could use to predict patient response to their MSC therapy. There were also concurrent preconference workshops on <a href="http://isct2018.com/isct-cba-cord-blood-series/" target="_blank">Cord Blood</a>, which featured several MSC talks, and <a href="http://isct2018.com/grp-workshop/" target="_blank">Global Regulatory Perspectives</a> and Standards. </span></div>
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<span style="font-family: "arial" , "helvetica" , sans-serif;">Another session that stood out was Progress in
the World of Standards for Cell and Gene Therapies. We discussed the need for
standards in an earlier <a href="http://roosterbio.blogspot.com/2014/04/regenerative-medicine-standards.html">blog post</a>. Jiwen Zhang, president
of the <a href="https://www.standardscoordinatingbody.org/">Standards Coordinating
Body</a><span class="MsoHyperlink">,</span> presented the group’s efforts to support
standards development, as well as training and educating the field on existing
and upcoming standards. This is important work as the gene, cell, and
regenerative medicine fields mature. RoosterBio looks forward to answering the <b style="mso-bidi-font-weight: normal;">SCB’s call for increased stakeholder
participation in standards development. </b><span style="mso-bidi-font-weight: normal;">You can download the Regenerative Medicine Standards Landscape State of the Industry Report from the SCB <a href="https://static1.squarespace.com/static/58a331b0db29d63c7fb64528/t/5ab254f9352f5362833b9cff/1521636606100/Landscape+Report_3-2-2018.pdf" target="_blank">here</a>.</span><o:p></o:p></span></div>
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<span style="font-family: "arial" , "helvetica" , sans-serif;">There were several sessions on <a href="https://www.bioinformant.com/types-stem-cell-derived-exosomes/" target="_blank">extracellular vesicles/exosomes</a> that highlighted the growing excitement in the field for a so-called “cell-free
therapy”. Sai King-Lim of <a href="https://www.a-star.edu.sg/" target="_blank">A-Star</a> gave a great talk summarizing the current
landscape called “MSC Exosomes and Small EVs: A Comparative Review”. Additionally,
the importance of a <b style="mso-bidi-font-weight: normal;">consistent
manufacturing process</b> was highlighted as numerous talks reported the impact
of manufacturing on extracellular vesicle phenotype and function. <o:p></o:p></span></div>
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<span style="font-family: "arial" , "helvetica" , sans-serif;">In addition to stem cells, the recent approvals
of <a href="https://www.cancer.gov/publications/dictionaries/cancer-terms/def/car-t-cell-therapy" target="_blank">CAR-T </a>immunotherapy drugs were much-discussed. One plenary session,
Reimagining Cancer Care and Delivering on the Promise of CAR-T Therapies,
featured a dynamic exchange between <a href="https://www.med.upenn.edu/cci/junelab/" target="_blank">Carl June of the University of Pennsylvania</a>,
<a href="https://www.reuters.com/article/us-novartis-pharmaceuticals/u-s-approves-novartis-cell-therapy-for-lymphoma-idUSKBN1I24GP" target="_blank">Pascal Touchon the of Novartis Oncology</a>, and <a href="https://www.kitepharma.com/about-us/" target="_blank">Diane Parks of Kite Pharma</a>. The
session discussed the development of CAR-T therapies in academia and the
translation to industry. Kite Pharma was a particularly interesting case as a
smaller, less well-known company that had to build trust with stakeholders like
doctors and patients as well as build an infrastructure to accomplish the
complicated logistics associated with autologous cell therapy before launching their
product Yescarta.<o:p></o:p></span></div>
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<span style="font-family: "arial" , "helvetica" , sans-serif;">Overall, the RoosterBio team had a great
conference connecting with current and future collaborators and customers as
well as other leaders in the field. The trip culminated with a Rooster Road
Trip after some bad weather and cancelled flights, but the team stuck it out
and ended up with a memorable drive back home to Frederick, Maryland.<o:p></o:p></span></div>
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<i style="mso-bidi-font-style: normal;"><span style="font-family: "arial" , "helvetica" , sans-serif;">Did you
attend ISCT? What were some of your takeaways?</span></i></div>
<br />Anonymousnoreply@blogger.com1tag:blogger.com,1999:blog-6181073719515632715.post-16125454416957317432017-06-29T12:49:00.002-04:002017-06-29T12:49:24.940-04:00Good Manufacturing Practice for Cell and Cell-Based Therapies: Facilities & Quality Control<div align="left" class="MsoNormal" style="line-height: 115%; margin-bottom: 0.0001pt;">
Novel cell and cell-based therapies require
stringent manufacturing, testing and oversight to ensure integrity, function,
and above all else, patient safety upon administration. Tissue-derived cellular products are
considered to be manufactured products and are regulated as such. Thus, you must ensure that your cell
manufacturing process is aligned to <a href="https://www.fda.gov/drugs/developmentapprovalprocess/manufacturing/ucm169105.htm">current
Good Manufacturing Practice requirements</a>.
(Note the “current”. This
means that these are evolving requirements, so you must stay up-to-date!) In the United States, human cells, tissue and
cellular- and tissue-based products (HCT/Ps) are regulated by the <a href="https://www.fda.gov/AboutFDA/CentersOffices/OfficeofMedicalProductsandTobacco/CBER/default.htm">Center
for Biologics Evaluation and Research</a> (CBER), a division of the <a href="http://www.fda.gov/">U.S. Food and Drug Administration</a> (FDA).<o:p></o:p></div>
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When manufacturing cell and cell-based
products, a production<b> facility</b>
under strict Quality Control must be used. Ideally, this facility includes the cell manufacturing suites, the storage space
for raw and finished product and any laboratory/testing areas. Thus, (1) facility design, access and maintenance, (2) equipment purchase, installation and operational qualification (I/OQ), use and maintenance and (3) raw material specifications,
purchase, use and storage must all be carefully controlled. For cell and cell-based therapies, terminal
sterilization of the final product is often not possible. As such, quality by design (QbD) is highly important in cell therapy, with stringent testing conducted
on the tissue Donor (our next blog post in this series will cover this topic)
to preclude risk of contamination at the source. In addition, facility and equipment standards
and monitoring must be instituted to ensure aseptic processing of cell and
cell-based products. To this end, <a href="http://cellculturedish.com/2015/04/closed-systems-in-biomanufacturing-offer-a-variety-of-benefits/">closed
systems</a> and <a href="http://www.contractpharma.com/issues/2016-03-01/view_features/single-use-technology-integral-to-advancing-biomanufacturing/11692">single-use
disposables</a> should be used whenever possible to minimize risk of
contamination. Therefore, a cGMP manufacturing facility must
include<a href="http://www.cleanairtechnology.com/cleanroom-classifications-class.php">
clean rooms</a> which control for temperature, humidity, pressure and air
particulates, preventing any contamination of manufactured product due to the environment, materials, human handlers and cross-contamination from other products manufactured in the same facility. As such, there should be
uni-directional flow of materials and people through these areas and personnel
must follow proper gowning procedures.
Facilities and equipment requirements are defined under US FDA <a href="https://www.accessdata.fda.gov/scripts/cdrh/cfdocs/cfcfr/CFRSearch.cfm?CFRPart=211">21CFR§211</a> and <a href="https://www.accessdata.fda.gov/scripts/cdrh/cfdocs/cfcfr/CFRSearch.cfm?CFRPart=1271" target="_blank">21CFR§1271</a>.<br />
<o:p></o:p></div>
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As mentioned above, stringent <b>Quality Control</b> <b>systems</b> must be in place to qualify all reagents and processes and
to institute <a href="https://en.wikipedia.org/wiki/Standard_operating_procedure">Standard
Operating Procedures</a> (SOPs) to ensure quality and consistency in
manufacturing processes and the end product.
<br />
<br />
<table align="center" cellpadding="0" cellspacing="0" class="tr-caption-container" style="margin-left: auto; margin-right: auto; text-align: center;"><tbody>
<tr><td style="text-align: center;"><a href="https://2.bp.blogspot.com/-xDe-28TGB3k/WVUvPBbYrOI/AAAAAAAAEoQ/Dyk71s2SKXodvOEVHwfM3um6FwhkV1ztACLcBGAs/s1600/Post%2B2%2Bgraphic.jpg" imageanchor="1" style="margin-left: auto; margin-right: auto;"><img border="0" data-original-height="1090" data-original-width="1600" height="435" src="https://2.bp.blogspot.com/-xDe-28TGB3k/WVUvPBbYrOI/AAAAAAAAEoQ/Dyk71s2SKXodvOEVHwfM3um6FwhkV1ztACLcBGAs/s640/Post%2B2%2Bgraphic.jpg" width="640" /></a></td></tr>
<tr><td class="tr-caption" style="text-align: center;">Facility and Quality Control considerations for cGMP cell manufacturing.</td></tr>
</tbody></table>
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<a name='more'></a>Such Quality Systems entail everything from personnel training, to proper
documentation of all aspects of your cell manufacturing process, to audits and
qualification of vendors for reagents and raw materials, to release of the end
product based on certain <a href="https://en.wikipedia.org/wiki/Critical_quality_attributes">Critical
Quality Attributes</a> (CQAs) and standard testing procedures (STPs) for testing
of your cell and cell-based products for these attributes. <b>Quality
Assurance</b> must also track any deviations and quickly implement
Corrective and Preventative Actions (CAPA) to rectify them and preclude their
repetition. To this end, Quality
Assurance works hand-in-hand with <b>Operations
and Manufacturing</b> to ensure standardized, well-documented procedures are
implemented, resulting in consistent, reproducible, safe end products. US FDA <a href="https://www.accessdata.fda.gov/scripts/cdrh/cfdocs/cfcfr/CFRSearch.cfm?CFRPart=1271" target="_blank">21CFR§1271</a> <a href="https://www.accessdata.fda.gov/scripts/cdrh/cfdocs/cfcfr/CFRSearch.cfm?fr=1271.160" target="_blank">Section 160</a> outlines the establishment and maintenance of a <b>Quality Program</b>. In addition, US FDA <a href="https://www.accessdata.fda.gov/scripts/cdrh/cfdocs/cfcfr/CFRSearch.cfm?CFRPart=211">21CFR§211</a> addresses Quality Control for cGMP manufacturing, and <a href="https://www.accessdata.fda.gov/scripts/cdrh/cfdocs/cfcfr/CFRSearch.cfm?fr=211.22">Section
22</a> specifically outlines the responsibilities of the Quality Control Unit.<br />
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The level of <b>processing and manufacturing oversight </b>that is needed for a
cellular therapy depends on a number of factors including, but not limited to,
the tissue source, the proliferation and differentiation potential of the
cells, the type of manipulations performed as a part of your cell
modification/manufacturing process, the intended use of the cells, the
anticipated method of cell integration into recipient tissues and organs, the
nature of the clinical trial and the number of patients who will be exposed to
the cellular product. Thus, requirements
for early-phase clinical trials may be vastly different, and less rigorous,
than those for later phase clinical trials, and subsequently for commercialization. Understanding these process and oversight
needs at each clinical stage is critical to successfully moving through the
clinical trial process towards rapid commercialization.<o:p></o:p></div>
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Anonymousnoreply@blogger.com0tag:blogger.com,1999:blog-6181073719515632715.post-64954074719978425092017-06-23T15:39:00.001-04:002017-06-26T11:55:43.038-04:00Demystifying Clinical Translation of Stem Cell-Based Therapies<div class="MsoNormal">
As our company matures and we continue to execute to our
mission of accelerating <a href="https://report.nih.gov/NIHfactsheets/ViewFactSheet.aspx?csid=62" target="_blank">Regenerative Medicine</a>, we find ourselves increasingly
involved in conversations around scalable cGMP cell manufacturing, qualification
of cell and cell-based therapies, and regulations around clinical translation
of <a href="https://www.roosterbio.com/pages/what-are-mscs" target="_blank">hMSCs</a> and hMSC-based therapeutics. While
the <a href="http://www.fda.gov/" target="_blank">US FDA</a> and other global regulatory agencies provide <a href="https://www.fda.gov/Drugs/GuidanceComplianceRegulatoryInformation/Guidances/ucm064971.htm" target="_blank">guidances</a> on the
regulations surrounding clinical translation of cellular products, it is
evident that there is still great uncertainty around rules, regulations,
processes, timelines and costs associated with clinical translation.</div>
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<i>How do you move from
pre-clinical to clinical studies?<o:p></o:p></i></div>
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<i>How must you
manufacture your cells?<o:p></o:p></i></div>
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<i>What data do you need
to present to the FDA to move to clinical trials?<o:p></o:p></i></div>
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<i>What sort of
characterization assays are needed?</i> <o:p></o:p></div>
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<i>What is the process
for engaging the FDA and moving to clinical trials?<o:p></o:p></i></div>
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<i>When do I need to do
all of this?!<o:p></o:p></i></div>
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<i><br /></i></div>
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<a href="https://1.bp.blogspot.com/-JYdwk9BXiPg/WU1t8E8-EZI/AAAAAAAAEl4/S80bmOgpxS8H0c5q4I8-p7CmqhdDI9KlQCLcBGAs/s1600/Post%2B1%2Bgraphic.jpg" imageanchor="1" style="margin-left: 1em; margin-right: 1em;"><img border="0" data-original-height="494" data-original-width="878" height="225" src="https://1.bp.blogspot.com/-JYdwk9BXiPg/WU1t8E8-EZI/AAAAAAAAEl4/S80bmOgpxS8H0c5q4I8-p7CmqhdDI9KlQCLcBGAs/s400/Post%2B1%2Bgraphic.jpg" width="400" /></a></div>
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These are just a few of the questions that we
commonly hear from our customers. In an
effort to facilitate our customers’ path to the clinic, we will be publishing
here a series of blog posts on clinical translation of hMSCs. These posts are meant to point you to the
relevant guidances and points to consider surrounding clinical translation of
hMSCs and are not meant to be the definitive authority on these matters. We recommend that you engage your Quality and
Regulatory teams (and/or consultants) early on in your Process Development
efforts to ensure that you are considering all regulations and guidelines in
developing a scalable, clinically- and commercially-relevant process and
product.<o:p></o:p></div>
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<br /></div>
<div align="left" class="MsoNormal" style="line-height: 115%; margin-bottom: 0.0001pt;">
<i>What
other questions do you have around clinical translation of hMSCs and hMSC-based
therapies?<o:p></o:p></i></div>
<br />
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Check back in next week for our first blog
post in this series.<o:p></o:p></div>
Anonymousnoreply@blogger.com0tag:blogger.com,1999:blog-6181073719515632715.post-37803513980687702392017-06-15T07:00:00.000-04:002017-06-15T07:00:39.440-04:00Opportunities for BioManufacturing Sciences to Accelerate Upscaled Mesenchymal Stem Cell Manufacturing TechnologiesContributed by: <span style="font-family: "times new roman"; font-size: 12pt; text-align: center;">Timothy R. Olsen, PhD - Sr. </span><span style="font-family: "times new roman"; font-size: 12pt; text-align: center;">Scientist, Process
and Product Development at RoosterBio Inc.</span><br />
<span style="font-family: "times new roman"; font-size: 12pt; text-align: center;"><br /></span>
<br />
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<a href="https://blogger.googleusercontent.com/img/b/R29vZ2xl/AVvXsEicOk-6MOXyn7Qse1tKeZ9VjuNZzgJkxkzBDf6TIVreBOWdeF2XqZAVmodXwjWQ0IS_pv2mEAvvYDQUTnHnXdmOQQZvdG9O4aLnOT2iftf8j3v9_vsulfM2mr7E04FS8BOGYX_o0fUINA/s1600/NIIMBL+Logo.png" imageanchor="1" style="clear: left; float: left; margin-bottom: 1em; margin-right: 1em;"><img border="0" data-original-height="144" data-original-width="350" height="163" src="https://blogger.googleusercontent.com/img/b/R29vZ2xl/AVvXsEicOk-6MOXyn7Qse1tKeZ9VjuNZzgJkxkzBDf6TIVreBOWdeF2XqZAVmodXwjWQ0IS_pv2mEAvvYDQUTnHnXdmOQQZvdG9O4aLnOT2iftf8j3v9_vsulfM2mr7E04FS8BOGYX_o0fUINA/s400/NIIMBL+Logo.png" width="400" /></a></div>
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After the National Institute for Standards and Technology (NIST) identified upcoming challenges in the U.S. biopharmaceutical
manufacturing landscape (see document <a href="http://cellmanufacturingusa.org/" target="_blank">here</a>), the <a href="http://www.niimbl.us/" target="_blank">National Institute for Innovation in Manufacturing Biopharmaceuticals (NIIMBL) </a>was formed to help solve these challenges through the formation of a public-private partnership between industry, government, academia, and non-profits. The goal of
NIIMBL is to accelerate Biopharmaceutical manufacturing innovation, support the
development of standards that enable more efficient and rapid manufacturing
capabilities, and educate and train a world-leading Biopharmaceutical manufacturing
workforce to maintain the United States’ global lead and competitiveness in
this industry. NIIMBL will be leveraging a $70 million
investment from NIST, with
at least $129 million more in funds from private partners. One of the great
aspects of this institute is that they are encouraging partnership between
large companies and smaller or medium size companies (called Small to Medium
Enterprises “SME”), which will be sure to bolster innovation and streamline
commercialization and implementation of new technologies. Kelvin Lee, the
NIIMBL Institute Director, did a fabulous job taking the lead on organizing the Consortium's first National Meeting, where members from the United States Congress, Directors
from the Food and Drug Administration, and many executive level industry
representatives were invited to speak about the importance of manufacturing
sciences and the current challenges we are facing as an industry. I had the
opportunity to represent RoosterBio as an SME at this inaugural NIIMBL National
Meeting, and I gave a talk in the “Rapid Fire” SME Innovation Showcase, as well
as presented some of our work on how we are radically shortening the
development timelines of Biopharmaceuticals that include a stem cell-derived
component.<o:p></o:p><br />
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<tr><td style="text-align: center;"><a href="https://blogger.googleusercontent.com/img/b/R29vZ2xl/AVvXsEh57n1k7HzXqIvjartn-D1dhXvijJ9UfjI1zgb7S4Klq4SZSXlLNakuUoQiySHJgGgLh4KIAy64NGhHjasPUg8aQeaAaIRd_YzKechDjES67a9vSzKPWdHcUDqRtlJet5RiOJJ9msDTYg/s1600/blue+microcarriers.PNG" imageanchor="1" style="clear: right; margin-bottom: 1em; margin-left: auto; margin-right: auto;"><img border="0" data-original-height="894" data-original-width="1188" height="240" src="https://blogger.googleusercontent.com/img/b/R29vZ2xl/AVvXsEh57n1k7HzXqIvjartn-D1dhXvijJ9UfjI1zgb7S4Klq4SZSXlLNakuUoQiySHJgGgLh4KIAy64NGhHjasPUg8aQeaAaIRd_YzKechDjES67a9vSzKPWdHcUDqRtlJet5RiOJJ9msDTYg/s320/blue+microcarriers.PNG" width="320" /></a></td></tr>
<tr><td class="tr-caption" style="text-align: center;">Confluent hMSCs on Solohill microcarriers. <br />
Image from RoosterBio Inc</td></tr>
</tbody></table>
Among
the many relevant talks given throughout the day, one specifically caught
my attention. In his talk titled “Key Process & Assay Challenges in Cell
Therapy Development,” <a href="https://www.linkedin.com/in/greg-russotti-0397686/" target="_blank">Greg Russotti</a> (Vice President, Technical Operations at
Celgene Cellular Therapeutics) laid out challenges in the upscaled
manufacturing of human mesenchymal stem cells (hMSCs). To meet the
pressing need for economical manufacturing of hMSCs at clinically- and commercially-
relevant scales, researchers have turned to single-use bioreactor systems that
have successfully been used to manufacture other biomolecules, such as
monocolonal antibodies which make up the lion’s share of <a href="http://labiotech.eu/top-10-best-selling-biologicals-blockbusters-2015/" target="_blank">blockbuster pharmaceuticals</a>.
However, unlike small molecule or large molecule production, cell therapy
products are living, breathing cells, which presents unique bioprocessing
constraints and challenges. Greg noted that there are technologies based on monoclonal
production that can expand hMSCs in large quantities, like using <a href="https://biopharmaceuticals.pall.com/en/webinars/accelerating-development-regenerative-therapies.html" target="_blank">3D microcarrier-based bioreactor systems</a>,
but there are still many manufacturing innovations required before these
manufacturing platforms can support a commercial cell therapy product.<span style="mso-spacerun: yes;"> </span></div>
<div class="MsoNormal" style="line-height: 150%; text-align: justify; text-indent: .5in;">
The <b><u>technology gaps</u></b> that he specifically
mentioned for upscaled hMSC manufacturing were <a href="http://www.bioprocessintl.com/upstream-processing/bioreactors/downstream-technology-landscape-for-large-scale-therapeutic-cell-processing-340981/" target="_blank">d</a><span style="font-size: 12pt; text-indent: 0.5in;"><a href="http://www.bioprocessintl.com/upstream-processing/bioreactors/downstream-technology-landscape-for-large-scale-therapeutic-cell-processing-340981/" target="_blank">ownstream processing technologies</a>, specifically the unit operations related to:</span><br />
<a name='more'></a></div>
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<o:p></o:p></div>
<div class="MsoListParagraphCxSpFirst" style="line-height: 150%; margin-left: 1.25in; mso-add-space: auto; mso-list: l0 level2 lfo1; text-align: justify; text-indent: -.25in;">
<!--[if !supportLists]--><span style="mso-bidi-font-family: "Times New Roman"; mso-fareast-font-family: "Times New Roman";"><span style="mso-list: Ignore;">a.<span style="font: 7.0pt "Times New Roman";"> </span></span></span>detaching
(or harvesting) the cells from microcarriers without damaging or changing the
cells in any way, <o:p></o:p></div>
<div class="MsoListParagraphCxSpMiddle" style="line-height: 150%; margin-left: 1.25in; mso-add-space: auto; mso-list: l0 level2 lfo1; text-align: justify; text-indent: -.25in;">
<!--[if !supportLists]--><span style="mso-bidi-font-family: "Times New Roman"; mso-fareast-font-family: "Times New Roman";"><span style="mso-list: Ignore;">b.<span style="font: 7.0pt "Times New Roman";"> </span></span></span>separating the detached cells from microcarriers
in suspension using an efficient, automated methodology, and <o:p></o:p></div>
<div class="MsoListParagraphCxSpLast" style="line-height: 150%; margin-left: 1.25in; mso-add-space: auto; mso-list: l0 level2 lfo1; text-align: justify; text-indent: -.25in;">
<!--[if !supportLists]--><span style="mso-bidi-font-family: "Times New Roman"; mso-fareast-font-family: "Times New Roman";"><span style="mso-list: Ignore;">c.<span style="font: 7.0pt "Times New Roman";">
</span></span></span><!--[endif]-->concentrating large volumes of bulk cell
solution in a timely manner (<5 hours) to maintain cell viability and functionality.<o:p></o:p></div>
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<br /></div>
<div class="MsoNormal" style="line-height: 150%; text-align: justify;">
I would like to
specifically address a couple of these challenges below: <o:p></o:p></div>
<div class="MsoNormal" style="line-height: 150%; text-align: justify; text-indent: .5in;">
The
harvest unit operation challenge mentioned above is due to the use of proteolytic
enzymes (i.e. trypsin) and agitation (i.e. shear forces) during cell dissociation, to dislodge cells from microcarriers, and is currently the industry standard.<span style="mso-spacerun: yes;"> </span>This unit operation must be done quickly and efficiently,
as over-trypsinization and long term exposure to high shear forces can be harmful
to the cells. To address some of limitations of conventional harvest enzymes, dissociation
reagents like TrypLE Select (Gibco-ThermoFisher), have been developed with
animal component-free materials, at cGMP grade and in a ready-to-use solution,
which will help to ease future regulatory burden. After the dissociation
enzymes have performed their function of detaching the cells from microcarriers, quenching the activity of the enzymes is critical for maintaining
viability of the bulk cell solution for further downstream processing. The
industry standard has been to use a solution of 2% fetal bovine serum in
phosphate buffered saline, but the presence of animal-derived components and variation
in efficacy of this quench solution pose challenges to its use in robust,
reproducible, cGMP-aligned manufacturing processes.. Thus, there is a
definitive need to develop a xeno-free alternative with <span style="mso-spacerun: yes;"> </span>validated inactivation of dissociation enzymes
to ensure optimal bioprocessing conditions.<o:p></o:p></div>
<div class="MsoNormal" style="line-height: 150%; text-align: justify; text-indent: .5in;">
Harvesting
of cells from microcarriers is then immediately followed by filtration of the
cell/microcarrier slurry through a porous mesh that traps the microcarriers,
while letting cells pass through. Dr. Russotti stressed that confirming
complete removal of the microcarriers from the cell therapy product is <b style="mso-bidi-font-weight: normal;"><u>absolutely critical</u></b> to ensuring
safety for the patient. At the small (0.1L to 5L) to medium (5L to 200L) scale bioreactor
size, conventional flow filtration technology has worked, but when the scale
increases to 200L and more, the volume of product to process can overwhelm most
systems. Upscaled technologies, like continuous flow centrifugation (kSep),
have been tested for cell suspensions but will require development and
validation for use with cell/microcarrier suspensions. A new and promising
technology innovation is the <span style="mso-spacerun: yes;"> </span>development
of <a href="http://csmedia2.corning.com/LifeSciences/media/pdf/poster_CLS-BP-PST-020_New_Microcarrier_Platform_BPI_2015.pdf" target="_blank">completely dissolvable microcarriers</a>,
which has the potential to obviate the cell/microcarrier separation unit
operation in the future, markedly streamlining and simplifying the downstream
processing of hMSCs expanded in microcarrier-based bioreactor systems. <b style="mso-bidi-font-weight: normal;"><o:p></o:p></b></div>
<div class="MsoNormal" style="line-height: 150%; text-align: justify; text-indent: .5in;">
While
there are several additional manufacturing challenges that will need to be
addressed prior to widespread commercial adoption of hMSC therapies, each of these
challenges creates opportunities to develop incrementally and radically innovative
products and processes as solutions. NIIMBL represents the perfect platform to
approach the challenges in upscaled cell therapy manufacturing through the
funding and support of proposals aimed at addressing these industry needs. Currently,
there are over 700 clinical trials (clinicaltrials.gov) using hMSCs, a number
which has more than doubled since 2014, and hMSCs <a href="https://www.blogger.com/null" style="mso-comment-date: 20170612T1544; mso-comment-reference: PB_1;">are poised to be the transistor and
microchip equivalent of tomorrow’s Regenerative Medicine technology products</a>.
These therapeutic cells are critical to many Regenerative Medicine applications, including cell and gene therapy, bioprinting, and tissue regeneration. The
many manufacturing science challenges like those outlined above create the
exact motivation for technology innovations with the development of which <span style="mso-spacerun: yes;"> </span>NIIMBL is tasked. <o:p></o:p></div>
<div class="MsoNormal" style="line-height: 150%; text-align: justify; text-indent: .5in;">
Given
the large patient populations for various target indications (stroke, Graft
Versus Host Disease, traumatic brain injury, cardiovascular disease) and dose
sizes of 10s to 100s of millions of cells per patient, <b style="mso-bidi-font-weight: normal;">upscaled manufacturing science technologies will play a critical role
in addressing the growing demand for significant volumes of pharmaceutical
quality hMSCs</b>. Advancements in Regenerative Medicine are fueling the need
for rapid
manufacturing science progress, especially with accelerated approval pathways
for RegenMed drugs being established around the world. RoosterBio, with our
disruptive approach to hMSC manufacturing and use, is uniquely poised to<span style="mso-spacerun: yes;"> </span>work alongside NIIMBL and other member
organizations to push forward <span style="mso-spacerun: yes;"> </span>hMSC-based
therapeutic platforms. We are excited to do our part in revolutionizing Biopharmaceutical manufacturing in the coming years.<o:p></o:p></div>
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</style>Jon Rowleyhttp://www.blogger.com/profile/10634518820328346758noreply@blogger.com0tag:blogger.com,1999:blog-6181073719515632715.post-39914297973626460502017-04-14T12:25:00.000-04:002017-04-14T14:22:25.367-04:00Orthopaedic Research Spotlight: ORS 2017 Annual Meeting<i><span style="font-family: "arial" , "helvetica" , sans-serif;">A guest blog post by RoosterBio Travel Award winner, <a href="http://www.orthomechlab.umd.edu/People/PS.html" target="_blank">Poonam Sharma</a>.</span></i><br />
<span style="font-family: "arial" , "helvetica" , sans-serif;"><i><br /></i>
</span><br />
<div class="MsoNormal">
<span style="font-family: "arial" , "helvetica" , sans-serif;">The annual
Orthopaedic Research Society meeting was an energetic and collaborative
conference attended by clinicians, industry professionals, and researchers. While
the attendees brought diverse perspectives to this meeting, the varied
presentation styles, such as short poster teasers, mid-length research talks,
and longer, broader spotlight oral presentations helped bring the audience
together in scientific discourse. With over 300 oral presentations and over 2200
poster presentations, the variety in presentation styles made <a href="http://www.ors.org/2017annualmeeting/" target="_blank">ORS 2017</a> an
engaging and dynamic conference to attend. <o:p></o:p></span></div>
<div class="MsoNormal">
<span style="font-family: "arial" , "helvetica" , sans-serif;"><br /></span></div>
<table cellpadding="0" cellspacing="0" class="tr-caption-container" style="float: right; margin-left: 1em; text-align: right;"><tbody>
<tr><td style="text-align: center;"><a href="https://2.bp.blogspot.com/-MuJgsf01010/WPD02hiam4I/AAAAAAAAEd8/yvmE1BryyEATqgKypPkulpGQgruaTHiqgCLcB/s1600/Sharma_Figure%2Bfor%2Bblog%2Bpost.jpg" imageanchor="1" style="clear: right; margin-bottom: 1em; margin-left: auto; margin-right: auto;"><span style="font-family: "arial" , "helvetica" , sans-serif;"><img border="0" height="320" src="https://2.bp.blogspot.com/-MuJgsf01010/WPD02hiam4I/AAAAAAAAEd8/yvmE1BryyEATqgKypPkulpGQgruaTHiqgCLcB/s320/Sharma_Figure%2Bfor%2Bblog%2Bpost.jpg" width="242" /></span></a></td></tr>
<tr><td class="tr-caption" style="text-align: center;"><span style="font-family: "arial" , "helvetica" , sans-serif; font-size: small;">Knockdown of vimentin may affect chondrogenic </span><br />
<span style="font-family: "arial" , "helvetica" , sans-serif; font-size: small;">extracellular matrix deposition in high density pellets. </span><br />
<span style="font-family: "arial" , "helvetica" , sans-serif; font-size: small;">shVim-vimentinknockdown, shLacZ-control.</span></td></tr>
</tbody></table>
<div class="MsoNormal">
<span style="font-family: "arial" , "helvetica" , sans-serif;">My
research in Dr. Adam H. Hsieh’s <a href="http://orthomechlab.umd.edu/" target="_blank">Orthopaedic Mechanobiology Lab</a> at the University of Maryland centers on the role
of vimentin intermediate filaments in governing <a href="https://www.roosterbio.com/pages/what-are-mscs" target="_blank">mesenchymal stem cell </a>(MSC) properties and behavior, including cellular deformability, adhesion, and
differentiation, specifically chondrogenesis. After knocking down the
expression of vimentin intermediate filaments in human MSCs using RNA interference, we
observe how a decrease in vimentin affects differentiation (abstract <a href="https://cdn.shopify.com/s/files/1/0515/3253/files/Sharma_RoosterBioTravelAward_ORS_2017_Abstract.pdf?4220194646026499655" target="_blank">here</a>). Preliminarily,
we’ve found that a decrease in vimentin did not affect adipogenesis or
osteogenesis, but may lead to a potential decrease in chondrogenic extracellular
matrix deposition, but this needs further exploration. <a href="https://www.roosterbio.com/" target="_blank">MSCs from RoosterBio</a>
have been singular in the progression of my graduate research. The fast growth
and consistency of the high quality MSCs have taken the bottleneck of MSC
growth out of the equation for my research. Further, using these MSCs and media
has dramatically decreased the labor, time, and resources needed to obtain the
cell numbers needed for conducting my experiments. <o:p></o:p></span></div>
<div class="MsoNormal">
<span style="font-family: "arial" , "helvetica" , sans-serif;"><br /></span></div>
<div class="MsoNormal">
<div class="separator" style="clear: both; text-align: center;">
<a href="https://4.bp.blogspot.com/-rVwIXelOVm8/WPD5a4QCwMI/AAAAAAAAEeA/jTjH3iO7ErkT9DOTh26g14X2HKWg5DJ2wCLcB/s1600/14231.jpeg" imageanchor="1" style="clear: left; float: left; margin-bottom: 1em; margin-right: 1em;"><img border="0" height="240" src="https://4.bp.blogspot.com/-rVwIXelOVm8/WPD5a4QCwMI/AAAAAAAAEeA/jTjH3iO7ErkT9DOTh26g14X2HKWg5DJ2wCLcB/s320/14231.jpeg" width="320" /></a></div>
<span style="font-family: "arial" , "helvetica" , sans-serif;">During this
conference, I was able to have in-depth conversations about my research as well
as exchange ideas and technical tips that will help strengthen my work.
Attending ORS allowed me to both present my research through a poster
presentation and network with both industry professionals and academic researchers.
As I will soon be taking the next step in my career, these interactions helped
me to start to home in on the types of opportunities that I would like to
pursue and how to prepare myself to excel. Further, attending professional
development seminars, such as one regarding the art of negotiation, helped me
identify techniques for further developing soft skills. <o:p></o:p></span></div>
<div class="MsoNormal">
<span style="font-family: "arial" , "helvetica" , sans-serif;">One of the
most engaging sessions in this conference was a really fun debate about the
related futures of regenerative medicine and orthopaedic implants – <i>Will Regenerative Medicine Make Orthopaedic Implants
Obsolete in Our Time?</i> It was captivating to hear the discussion about two research
and clinical areas that continue to intersect and diverge. Also, the keynote by
<a href="http://rna.berkeley.edu/" target="_blank">Dr. Jennifer Doudna </a>summarizing the <a href="http://gizmodo.com/everything-you-need-to-know-about-crispr-the-new-tool-1702114381" target="_blank">CRISPR technology</a> that she helped develop
was a great overview and the brief discussion about the ethics of gene therapy
was thought-provoking. <o:p></o:p></span></div>
<div class="MsoNormal">
<span style="font-family: "arial" , "helvetica" , sans-serif;"><br /></span></div>
<div class="MsoNormal">
<span style="font-family: "arial" , "helvetica" , sans-serif;">The
research presentations and broader spotlight sessions gave me a great overview
of the latest research in my interest areas of regenerative medicine, tissue
engineering, cell therapies, and biomaterials. Many of the oral presentations I
attended focused on bettering the design of tissue engineered scaffolds. Here
are just a few of the research presentations that inspired me:<o:p></o:p></span></div>
<div class="MsoNormal">
<span style="font-family: "arial" , "helvetica" , sans-serif;"></span></div>
<a name='more'></a><span style="font-family: "arial" , "helvetica" , sans-serif;"><span style="font-family: "times" , "times new roman" , serif;"><br /></span>
<span style="font-family: "arial" , "helvetica" , sans-serif;"><span style="font-family: "times" , "times new roman" , serif;"><br /></span>
</span></span><br />
<div class="MsoNormal">
<b><span style="font-family: "arial" , "helvetica" , sans-serif;">Development of a Continuous
Dual-Gradient Peptide Scaffold for Directed Osteochondral Spatial
Differentiation<o:p></o:p></span></b></div>
<div class="MsoListParagraphCxSpFirst" style="mso-list: l2 level1 lfo1; text-indent: -.25in;">
<!--[if !supportLists]--><span style="font-family: "arial" , "helvetica" , sans-serif;"><span style="line-height: 115%;">·<span style="font-stretch: normal; line-height: normal;">
</span></span><!--[endif]--><span style="font-family: "times new roman" , serif;">Dr.
Jennifer L. Puetzer, <a href="http://www.stevensgroup.org/" target="_blank">Stevens Lab, Imperial College London </a><o:p></o:p></span></span></div>
<div class="MsoListParagraphCxSpMiddle" style="mso-list: l2 level1 lfo1; text-indent: -.25in;">
<!--[if !supportLists]--><span style="font-family: "arial" , "helvetica" , sans-serif;"><span style="line-height: 115%;">·<span style="font-stretch: normal; line-height: normal;">
</span></span><!--[endif]--><span style="font-family: "times new roman" , serif;">Dr.
Puetzer presented a sequentially electrospun polycaprolactone scaffold with
opposing gradients of two peptides in a single scaffold for osteochondral
repair. MSCs seeded on the glycosaminoglycan-binding
peptide side of the scaffold showed chondrogenic differentiation while those
seeded on the mineral nucleating peptide side showed osteogenic differentiation
without any exogenous growth factors! <o:p></o:p></span></span></div>
<div class="MsoListParagraphCxSpLast" style="mso-list: l2 level1 lfo1; text-indent: -.25in;">
<!--[if !supportLists]--><span style="font-family: "arial" , "helvetica" , sans-serif;"><span style="line-height: 115%;">·<span style="font-stretch: normal; line-height: normal;">
</span></span><!--[endif]--><span style="font-family: "times new roman" , serif;">Learn
more about their dual gradient peptide research <a href="http://onlinelibrary.wiley.com/doi/10.1002/adhm.201600699/abstract" target="_blank">here</a>.</span></span></div>
<div class="MsoListParagraphCxSpLast" style="mso-list: l2 level1 lfo1; text-indent: -.25in;">
<span style="font-family: "arial" , "helvetica" , sans-serif;"><br /></span></div>
<div class="MsoNormal">
<b><span style="font-family: "arial" , "helvetica" , sans-serif;">A Bi-layered Scaffold Derived from
Decellularized Growth Plate and Articular Cartilage Extracellular Matrix for
Osteochondral Defect Repair<o:p></o:p></span></b></div>
<div class="MsoListParagraphCxSpFirst" style="mso-list: l0 level1 lfo2; text-indent: -.25in;">
<!--[if !supportLists]--><span style="font-family: "arial" , "helvetica" , sans-serif;"><span style="line-height: 115%;">·<span style="font-stretch: normal; line-height: normal;">
</span></span><!--[endif]--><span style="font-family: "times new roman" , serif;">Dr.
Grainne Cunniffe, <a href="http://www.mee.tcd.ie/regenerative/People/DKelly" target="_blank">Kelly Lab, Trinity College Dublin</a><o:p></o:p></span></span></div>
<div class="MsoListParagraphCxSpMiddle" style="mso-list: l0 level1 lfo2; text-indent: -.25in;">
<!--[if !supportLists]--><span style="font-family: "arial" , "helvetica" , sans-serif;"><span style="line-height: 115%;">·<span style="font-stretch: normal; line-height: normal;">
</span></span><!--[endif]--><span style="font-family: "times new roman" , serif;">Dr.
Cunniffe presented a bi-layered scaffold for osteochondral repair using
decellularized and homogenized growth plate and articular cartilage
extracellular matrices. <i>In vitro</i>,
MSCs seeded onto the scaffold differentiated in response to the different
extracellular matrix cues, with more glycosaminoglycans found on the cartilage-derived
matrix and calcification found on the growth plate-derived matrix. In an <i>in vivo</i> goat osteochondral defect model,
hyaline cartilage and endochondral ossification were observed by 12 months. <o:p></o:p></span></span></div>
<div class="MsoListParagraphCxSpLast" style="mso-list: l0 level1 lfo2; text-indent: -.25in;">
<!--[if !supportLists]--><span style="font-family: "arial" , "helvetica" , sans-serif;"><span style="line-height: 115%;">·<span style="font-stretch: normal; line-height: normal;">
</span></span><!--[endif]--><span style="font-family: "times new roman" , serif;">Learn
more about their extracellular matrix-derived scaffolds <a href="https://www.ncbi.nlm.nih.gov/pubmed/28197989" target="_blank">here</a>.<o:p></o:p></span></span></div>
<div class="MsoNormal">
<span style="font-family: "arial" , "helvetica" , sans-serif;"><br />
<b>Mesenchymal Stem Cells Expressing the
Cell Adhesion Molecule CD146 Present Increased Homing Towards Degenerative
Intervertebral Discs - an Organ Culture Study<o:p></o:p></b></span></div>
<div class="MsoListParagraphCxSpFirst" style="mso-list: l1 level1 lfo3; text-indent: -.25in;">
<!--[if !supportLists]--><span style="font-family: "arial" , "helvetica" , sans-serif;"><span style="line-height: 115%;">·<span style="font-stretch: normal; line-height: normal;">
</span></span><!--[endif]--><span style="font-family: "times new roman" , serif;">Sebastian
Wangler, <a href="https://www.aofoundation.org/Structure/research/exploratory-applied-research/research-institute/research-institute-services/musculoskeletal-regeneration/Pages/Sibylle-Grad.aspx" target="_blank">Grad Lab, AO Research Institute</a></span></span></div>
<div class="MsoListParagraphCxSpMiddle" style="mso-list: l1 level1 lfo3; text-indent: -.25in;">
<!--[if !supportLists]--><span style="font-family: "arial" , "helvetica" , sans-serif;"><span style="line-height: 115%;">·<span style="font-stretch: normal; line-height: normal;">
</span></span><!--[endif]--><span style="font-family: "times new roman" , serif;">Sebastian
Wangler presented a series of studies to characterize MSC homing to
degenerative intervertebral discs. MSCs migrating toward both a degenerative
disc chemoattractant and conditioned media from an induced degenerative disc
model expressed higher levels of the marker CD146 compared to controls.
Further, in an induced degenerative disc organ study, CD146+ MSCs showed
increased migration into the discs compared to CD146- MSCs. <b><o:p></o:p></b></span></span></div>
<div class="MsoListParagraphCxSpLast" style="mso-list: l1 level1 lfo3; text-indent: -.25in;">
<!--[if !supportLists]--><span style="font-family: "arial" , "helvetica" , sans-serif;"><span style="line-height: 115%;">·<span style="font-stretch: normal; line-height: normal;">
</span></span><!--[endif]--><span style="font-family: "times new roman" , serif;">Learn
more about MSC homing to chemoattractant cues from degenerative discs <a href="http://onlinelibrary.wiley.com/doi/10.1002/jor.23326/full" target="_blank">here</a>.</span></span></div>
<div class="MsoListParagraphCxSpLast" style="mso-list: l1 level1 lfo3; text-indent: -.25in;">
<span style="font-family: "arial" , "helvetica" , sans-serif;"><br /></span></div>
<span style="font-family: "arial" , "helvetica" , sans-serif;"><span style="font-family: "times" , "times new roman" , serif;"><br /></span>
</span><br />
<div class="MsoNormal">
<span style="font-family: "times new roman" , serif;"><span style="font-family: "arial" , "helvetica" , sans-serif;">This
year’s ORS annual meeting was really engaging and was excellent for fostering
discussion about the latest research in orthopaedics, professional development,
and networking. I enjoyed presenting my research at ORS and am thankful to
RoosterBio for their high-quality MSCs, media systems, and service as well as
their support toward my attendance at ORS 2017! </span><span style="font-family: "times new roman" , serif;"><o:p></o:p></span></span></div>
Anonymousnoreply@blogger.com0tag:blogger.com,1999:blog-6181073719515632715.post-30834421261683244622017-01-03T11:42:00.000-05:002017-01-03T11:45:21.644-05:00Guest Blog Post: The Ambitious Future of Tissue Engineering<h4 style="text-align: justify; text-indent: .25in;">
<i>Bagrat Grigoryan and <a href="http://bioengineering.rice.edu/faculty/Jordan_Miller.aspx" target="_blank">Jordan Miller</a>, <a href="http://millerlab.rice.edu/" target="_blank">Physiologic Systems Engineering & Advanced Materials Lab </a>at Rice University.</i></h4>
<div class="MsoNormal" style="text-align: justify; text-indent: .25in;">
<br /></div>
<div class="MsoNormal" style="text-align: justify; text-indent: .25in;">
Over the last
several decades, various advances in <a href="https://en.wikipedia.org/wiki/Tissue_engineering" target="_blank">tissue engineering</a> have allowed for the
not so distant possibility of replacing, repairing, or regenerating injured
tissues<!--[if supportFields]><span style='mso-element:field-begin;mso-field-lock:
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style='mso-element:field-end'></span><![endif]-->. Significant progress has been
made in understanding cellular biology as well as pathophysiology and healthy
states of tissues. Additionally, a suite of diverse <a href="https://biofabdegree.net/what-is-biofabrication-2/" target="_blank">biofabrication</a> technologies
and <a href="https://en.wikipedia.org/wiki/Biomaterial" target="_blank">biomaterials</a> has been conceived, enabling fabrication of complex 3D tissues
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past several decades, there has been an ever-increasing demand for organ
transplants. However, there is a severe shortage of donor organs, and as a
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using biomaterial scaffolds and a person's own cells. Although the realization
of this solution has been limited, the development of new biofabrication
approaches has made it more realistic. This review provides an overview of
natural and synthetic biomaterials that have been used for organ/tissue
development. It then discusses past and current biofabrication techniques, with
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"page" : "247-76", "title" : "3D Biofabrication
Strategies for Tissue Engineering and Regenerative Medicine.",
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}<span style='mso-element:field-separator'></span><![endif]--><sup>2</sup><!--[if supportFields]><span
style='mso-element:field-end'></span><![endif]-->. However, the field of tissue
engineering still has unresolved questions involving choice of fabrication
technique, biomaterial, cellular niche, or even cell type when designing a
synthetic tissue.<o:p></o:p></div>
<div class="MsoNormal" style="text-align: justify; text-indent: .25in;">
<br /></div>
<div class="MsoNormal" style="text-align: justify; text-indent: .25in;">
While different
fabrication techniques and biomaterials have been explored in fabricating
tissues <i>in vitro</i>, the use of stem
cells in engineered tissues is ubiquitous. Not surprisingly so, as biologists
continually demonstrate novel ways of directing different lineage commitment of
stem cells and further unlocking their vast regenerative potential<!--[if supportFields]><span
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extremely promising therapeutic agent for tissue regeneration. Studies in
animal models of myocardial infarction have demonstrated the ability of
transplanted MSCs to engraft and differentiate into cardiomyocytes and
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array of paracrine factors. Together, these properties can be harnessed to both
prevent and reverse remodeling in the ischemically injured ventricle. In
proof-of-concept and phase I clinical trials, MSC therapy improved left
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}<span style='mso-element:field-separator'></span><![endif]--><sup>3</sup><!--[if supportFields]><span
style='mso-element:field-end'></span><![endif]-->. Indeed, stem cell banks have
emerged to cryogenically store a patient’s own cells as the therapeutic
potential of stem cells is being positively demonstrated in multiple clinical
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time of collection, compromising their efficacy. Stem cell banking offers the
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style='mso-element:field-end'></span><![endif]-->. With <a href="https://www.ncbi.nlm.nih.gov/pubmed/26423725" target="_blank">over 450 mesenchymal stem cell (MSC)-based trials </a>alone currently ongoing or completed, the
<a href="http://www.roosterbio.com/pages/what-are-mscs" target="_blank">regenerative and immunoregulatory properties of MSCs</a> are constantly being
exploited to improve the quality of human life<!--[if supportFields]><span
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their regenerative properties and ability to differentiate into diverse cell
lineages, have generated great interest among researchers whose work has
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Currently the most commonly used adult stem cells in regenerative medicine,
MSCs can be isolated from several tissues, exhibit a strong capacity for
replication in vitro, and can differentiate into osteoblasts, chondrocytes, and
adipocytes. However, heterogeneous procedures for isolating and cultivating
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cells. In particular, we analyze clinical trials using MSCs for representative
diseases, including hematological disease, graft-versus-host disease, organ transplantation,
diabetes, inflammatory diseases, and diseases in the liver, kidney, and lung,
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style='mso-element:field-end'></span><![endif]-->.<o:p></o:p></div>
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<br /></div>
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Due to the <a href="http://www.roosterbio.com/pages/what-are-mscs" target="_blank">immense therapeutic potential of MSCs</a>, there is a need to <a href="http://roosterbio.blogspot.com/2014/02/current-bottlenecks-in-msc-research.html" target="_blank">rapidly and reproducibly grow a vast amount of MSCs</a> for clinical and research purposes. Although MSCs were
identified and isolated from bone marrow more than 40 years ago, we still have
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animal models of myocardial infarction have demonstrated the ability of
transplanted MSCs to engraft and differentiate into cardiomyocytes and
vasculature cells, recruit endogenous cardiac stem cells, and secrete a wide
array of paracrine factors. Together, these properties can be harnessed to both
prevent and reverse remodeling in the ischemically injured ventricle. In
proof-of-concept and phase I clinical trials, MSC therapy improved left
ventricular function, induced reverse remodeling, and decreased scar size. This
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<a name='more'></a>Additionally, even though
the first MSC clinical trial occurred in 1995, we still do not have a complete
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including definition of the niche, and identification of signals regulating
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produced invaluable tools for the development of rational cell therapy protocols
that have yielded positive results in preclinical models of genetic and
acquired diseases and, in several cases, have entered clinical experimentation
with positive outcome. Adult mesenchymal stem cells (MSCs) are nonhematopoietic
cells with multilineage potential to differentiate into various tissues of
mesodermal origin. They can be isolated from bone marrow and other tissues and
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style='mso-element:field-end'></span><![endif]-->. Large expansion of MSCs for
research purposes is especially desirable for fabrication of synthetic tissues <i>in vitro</i>, especially as many available
technologies are demonstrating the ability to generate tissues in the micro- to
milli-liter volume scale<!--[if supportFields]><span style='mso-element:field-begin;
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style='mso-element:field-end'></span><![endif]-->. Additionally, as the cost of
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scalability of tissue fabrication is growing. Rapid expansion of MSCs will
allow for more experimentation <i>in vitro</i>,
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clinical work is considered. <o:p></o:p><br />
<br />
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Rapid expansion
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optimize different <a href="http://www.nature.com/nmeth/journal/v13/n1/full/nmeth.3702.html" target="_blank">labeling strategies</a> of hMSCs with fluorescent reports. We
have previously demonstrated stable transduction of RoosterBio hMSCs with a
second-generation lentivirus. We have achieved dual-labeling of hMSCs
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style='mso-element:field-end'></span><![endif]--> (Fig. 1A). Compared to cell
staining, constitutive labeling allows us to monitor cell behavior transiently,
in a non-destructive manner, throughout our studies. Dual-labelling of hMSCs
has been most useful in <a href="https://www.ncbi.nlm.nih.gov/pmc/articles/PMC4032528/" target="_blank">co-culture studies</a> where cell-cell interaction and morphological
and spatial changes can be easily monitored non-destructively at multiple time
points. Additionally, dual-labeling allows us to identify cytoplasmic and
nuclear regions in aggregates that are encapsulated in a 3D context (Fig. 1B). We
have also shown osteogenic (bone) differentiation of unlabeled hMSCs seeded on <a href="https://en.wikipedia.org/wiki/3D_printing" target="_blank">3D printed</a> PCL platforms<!--[if supportFields]><span style='mso-element:field-begin;
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<br /></div>
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As we look
toward the future, we believe that fundamental engineering and biological
concepts should further fuse to maximize the translational capacity of stem
cells. The challenge tissue engineering poses is ambitious, however --
developing more physiologically relevant <i>in
vitro</i> models by incorporating approaches from both fields will progress
medicine and drastically improve the quality of human life. Additionally, akin
to stem cell banking for clinical use, rapid expansion and copious
cryopreservation of a batch of stem cells for research use (i.e. a thaw-and-use
stem cell product, such as <a href="http://www.prweb.com/releases/2015/10/prweb13027227.htm" target="_blank">RoosterRTP</a>) will allow researchers the ability to perform
experiments faster, with more consistency and reliability, as studies can be
performed with cells right out of thaw.<o:p></o:p></div>
<div class="MsoNormal" style="text-align: justify;">
<br /></div>
<div class="MsoNormal" style="text-align: justify;">
<b>References<o:p></o:p></b></div>
<div class="MsoNormal" style="line-height: normal; margin-left: 32.0pt; mso-layout-grid-align: none; mso-pagination: none; text-autospace: none; text-indent: -32.0pt;">
<!--[if supportFields]><span
style='mso-element:field-begin;mso-field-lock:yes'></span>ADDIN Mendeley
Bibliography CSL_BIBLIOGRAPHY <span style='mso-element:field-separator'></span><![endif]-->1. Khademhosseini, A. & Langer, R. <a href="http://www.nature.com/nprot/journal/v11/n10/full/nprot.2016.123.html" target="_blank">A decade of progress in tissue engineering.</a> <i>Nat. Protoc.</i> <b>11,</b>
1775–1781 (2016).<o:p></o:p></div>
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2. Bajaj, P., Schweller, R. M.,
Khademhosseini, A., West, J. L. & Bashir, R. 3<a href="https://www.ncbi.nlm.nih.gov/pubmed/24905875" target="_blank">D Biofabrication Strategies for Tissue Engineering and Regenerative Medicine.</a> <i>Annu. Rev. Biomed. Eng.</i>
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5. Squillaro, T., Peluso, G. &
Galderisi, U. <a href="https://www.researchgate.net/publication/282361576_Clinical_Trials_With_Mesenchymal_Stem_Cells_An_Update" target="_blank">Clinical Trials with Mesenchymal Stem Cells: An Update.</a> <i>Cell
Transplant.</i> 1–53 (2015). doi:10.3727/096368915X689622<o:p></o:p></div>
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7. Kinstlinger, I. S. & Miller, J.
<a href="http://pubs.rsc.org/en/content/articlelanding/2016/lc/c6lc00193a#!divAbstract" target="_blank">3D-printed Fluidic Networks as Vasculature for Engineered Tissue.</a> <i>Lab Chip</i>
(2016). doi:10.1039/C6LC00193A<o:p></o:p></div>
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<a href="http://pubs.rsc.org/en/Content/ArticleLanding/2016/RA/C5RA26022A#!divAbstract" target="_blank">Ultrahigh-throughput Generation and Characterization of Cellular Aggregates in Laser-ablated Microwells of Poly(dimethylsiloxane). </a><i>RSC Adv.</i> <b>6,</b>
8980–8991 (2016).<o:p></o:p></div>
<div class="MsoNormal" style="text-align: justify; text-indent: .25in;">
<!--[if supportFields]><span style='font-size:11.0pt;line-height:107%;
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<div class="MsoNormal" style="line-height: normal; margin-left: 32.0pt; mso-layout-grid-align: none; mso-pagination: none; text-autospace: none; text-indent: -32.0pt;">
9.<span style="mso-tab-count: 1;"> </span>Kinstlinger, I. S. <i>et al.</i>
<a href="http://journals.plos.org/plosone/article?id=10.1371/journal.pone.0147399" target="_blank">Open-Source Selective Laser Sintering (OpenSLS) of Nylon and Biocompatible Polycaprolactone.</a> <i>PLoS One</i> <b>11,</b> e0147399 (2016).<o:p></o:p></div>
Anonymousnoreply@blogger.com0tag:blogger.com,1999:blog-6181073719515632715.post-60379400113722492712016-06-16T10:41:00.000-04:002016-06-16T10:41:02.541-04:00At the Cutting Edge of 3D Bioprinting: WBC 2016 Round Up Part II<i>A guest blog post by RoosterBio Travel Award winner, <a href="http://millerlab.rice.edu/members/" target="_blank">Ian Kinstlinger</a>.</i><br />
<br />
<table align="center" cellpadding="0" cellspacing="0" class="tr-caption-container" style="float: left; margin-right: 1em; text-align: left;"><tbody>
<tr><td style="text-align: center;"><a href="https://3.bp.blogspot.com/-MGHRkMJUlA4/V2K5OXhdsPI/AAAAAAAAD7U/_jV7biU05n0QXMbj5rQk8zyI9h5zJyx0QCLcB/s1600/Kinstlinger1.jpg" imageanchor="1" style="clear: right; margin-bottom: 1em; margin-left: auto; margin-right: auto;"><img border="0" height="320" src="https://3.bp.blogspot.com/-MGHRkMJUlA4/V2K5OXhdsPI/AAAAAAAAD7U/_jV7biU05n0QXMbj5rQk8zyI9h5zJyx0QCLcB/s320/Kinstlinger1.jpg" width="240" /></a></td></tr>
<tr><td class="tr-caption" style="text-align: center;"><div align="center" class="MsoNormal">
<span style="font-size: 10.0pt; line-height: 107%; mso-bidi-font-size: 11.0pt;">Presenting
on Open-source Selective Laser </span></div>
<div align="center" class="MsoNormal">
<span style="font-size: 10.0pt; line-height: 107%; mso-bidi-font-size: 11.0pt;">Sintering </span><span style="font-size: 10pt; line-height: 107%;">at WBC2016!</span></div>
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international community of biomaterials scientists and engineers has convened
every four years to discuss the cutting edge of biomaterials research. This
year’s 10<sup>th</sup> World Biomaterials Congress (WBC) brought us to lovely
Montreal, Canada for a stimulating week of workshops, talks, posters, and
social activities. I was honored to present my work from the <a href="http://millerlab.rice.edu/">Miller Lab</a> at Rice University in both a podium
talk and a poster session. <o:p></o:p></div>
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<span
style='font-size:10.0pt;mso-bidi-font-size:11.0pt;line-height:107%'>Presenting
on Open-source Selective Laser Sintering at WBC2016!<o:p></o:p></span></p>
</div>
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</v:shape><![endif]--><!--[if !vml]--><!--[endif]-->Our lab is broadly interested in
developing strategies to construct vascular networks within engineered tissues.
In my research, I have developed a platform technology which uses 3D printed
carbohydrates as templates around which cells and biomaterials can be
assembled. Dissolving the sugar away gives you an engineered tissue with
perfusable channels; we believe that these constructs will be useful for
understanding the mass transport requirements and emergent properties of
engineered living tissue. <o:p></o:p></div>
<div class="MsoNormal">
<br /></div>
<table cellpadding="0" cellspacing="0" class="tr-caption-container" style="float: right; margin-left: 1em; text-align: right;"><tbody>
<tr><td style="text-align: center;"><a href="https://2.bp.blogspot.com/-kClOjwlYJ1I/V2K5az0yFuI/AAAAAAAAD7c/x7rquPrSHIo0B3SXjWUB_4aEUPfBDRC4gCLcB/s1600/Kinstlinger2.jpg" imageanchor="1" style="clear: left; margin-bottom: 1em; margin-left: auto; margin-right: auto;"><img border="0" height="232" src="https://2.bp.blogspot.com/-kClOjwlYJ1I/V2K5az0yFuI/AAAAAAAAD7c/x7rquPrSHIo0B3SXjWUB_4aEUPfBDRC4gCLcB/s320/Kinstlinger2.jpg" width="320" /></a></td></tr>
<tr><td class="tr-caption" style="text-align: center;"><div align="center" class="MsoNormal">
<span style="font-size: 10.0pt; line-height: 107%; mso-bidi-font-size: 11.0pt;">An overview
of one method our lab has introduced to create </span></div>
<div align="center" class="MsoNormal">
<span style="font-size: 10.0pt; line-height: 107%; mso-bidi-font-size: 11.0pt;">embedded vascular networks in
biomaterials.<o:p></o:p></span></div>
</td></tr>
</tbody></table>
<div class="MsoNormal">
I used my poster to
spread the word about our lab’s <a href="https://github.com/millerlabftw/opensls">Open-source Selective Laser
Sintering</a> technology and my podium talk to describe how we’ve adapted this
system to perform laser-based 3D printing of carbohydrate materials. I was
thrilled to have a large audience for my talk, followed by several insightful
questions. My poster also received a steady stream of visitors, many of whom
are involved in the open-source hardware community and were eager to talk about
hardware hacking for biomaterials. That work was actually <a href="http://journals.plos.org/plosone/article?id=10.1371/journal.pone.0147399">published
earlier this year</a> – and RoosterBio hMSCs were absolutely central. Their
high quality and robust differentiation response made characterizing
biocompatibility of materials quite straightforward.</div>
<div class="MsoNormal">
<o:p></o:p></div>
<div class="MsoNormal">
<br /></div>
<div class="MsoNormal">
A couple of key presentations stood out at WBC 2016:<o:p></o:p></div>
<div class="MsoNormal">
<u><br /></u></div>
<div class="MsoNormal">
<u>Nano- and Micro-fabricated Hydrogels for Regenerative
Engineering</u></div>
<div class="MsoNormal">
</div>
<ul>
<li><span style="text-indent: -0.25in;">Dr. Ali Khademhosseini, </span><a href="http://www.tissueeng.net/lab/" style="text-indent: -0.25in;">Khademhosseini Lab</a><span style="text-indent: -0.25in;">, Harvard University</span></li>
<li><span style="font-family: Symbol; text-indent: -0.25in;"><span style="font-family: 'Times New Roman'; font-size: 7pt; font-stretch: normal;"> </span></span><span style="text-indent: -0.25in;">Dr. Khademhosseini gave an illuminating keynote
on the many angles from which his lab is using bioprinting technologies to
fabricate functional biological structures. He is also emerging as a leader in
the field of integrated organ-on-chip drug screening platforms.</span></li>
</ul>
<br />
<div class="MsoNormal" style="background: white; line-height: 15.0pt; margin-bottom: .0001pt; margin-bottom: 0in; margin-left: 0in; margin-right: -11.25pt; margin-top: 0in;">
<u>Injection
of Dual-Crosslinking Hydrogels to Limit Infarct Induced Left Ventricular
Remodeling</u></div>
<div class="MsoNormal" style="background: white; line-height: 15.0pt; margin-bottom: .0001pt; margin-bottom: 0in; margin-left: 0in; margin-right: -11.25pt; margin-top: 0in;">
</div>
<ul>
<li><span style="background-color: transparent; line-height: 15pt; text-indent: -0.25in;">Dr. Jason Burdick, </span><a href="http://www.seas.upenn.edu/~burdick2/" style="background-color: transparent; line-height: 15pt; text-indent: -0.25in;">Polymeric Biomaterials Laboratory</a><span style="background-color: transparent; line-height: 15pt; text-indent: -0.25in;">,
University of Pennsylvania</span></li>
<li><span style="background-color: transparent; font-family: Symbol; line-height: 15pt; text-indent: -0.25in;"><span style="font-family: 'Times New Roman'; font-size: 7pt; font-stretch: normal;"> </span></span><span style="background-color: transparent; line-height: 15pt; text-indent: -0.25in;">The Burdick lab has developed an innovative
class of supramolecular biomaterials specifically targeted for 3D printing
applications. The gels are shear-thinning due to their non-covalent crosslinks,
and thus are amenable to extrusion printing. These materials are also useful as
injectables for reducing left ventricular remodeling after heart attack.</span></li>
</ul>
<br />
<div class="MsoNormal">
<u>Photoreversible
patterning of hydrogel biomaterials with site-specifically-modified proteins</u></div>
<div class="MsoNormal">
</div>
<ul>
<li><span style="text-indent: -0.25in;">Dr. Cole DeForest,</span><span style="text-indent: -0.25in;"> </span><a href="http://www.coledeforest.com/" style="text-indent: -0.25in;">DeForest
Research Group</a><span style="text-indent: -0.25in;">, University of Washington</span></li>
<li><span style="font-family: Symbol; text-indent: -0.25in;"><span style="font-family: 'Times New Roman'; font-size: 7pt; font-stretch: normal;"> </span></span><span style="text-indent: -0.25in;">Much like our lab is interested in patterning
biomaterial architecture via 3D printing, the DeForest group is patterning
functional proteins into materials through some very clever photochemistries.
Their techniques give them spatiotemporal control over the incorporation of
various full proteins into synthetic hydrogels.</span></li>
</ul>
<br />
<div class="MsoNormal">
It was tremendously exciting to see so many investigators
working on 3D printing of biomaterials. I counted at least seven sessions
devoted to the topic and was also impressed by the low-cost <a href="file:///C:/Users/Betty/Downloads/biobots.io">printers</a> and <a href="file:///C:/Users/Betty/Downloads/cellink.edu">inks</a> now hitting the
market, including RoosterBio’s new <a href="http://www.roosterbio.com/collections/cells-1/products/roosterrtp-hbm-50m-msc-006">ready-to-print
hMSC products</a>. The diverse hardware and materials that have been introduced
in the past few years are already transforming the field! It will be very
interesting to see in the coming years whether these new techniques give way to
novel insights into cell and tissue function <i>in vitro</i>, as many groups are currently promising.</div>
<div class="MsoNormal">
<br /></div>
<div class="MsoNormal">
It is also not yet clear whether the same groups who are
mastering the materials and fabrication technology have the resources and
expertise to analyze complex biological phenomena in their printed structures.
A greater level of collaboration between biologists and materials/fabrication
engineers may be necessary in the future to make progress in this area. I am
going to end with shameless plug for my <a href="http://pubs.rsc.org/en/content/articlelanding/2016/lc/c6lc00193a">recent
review article</a> in Lab on a Chip which discusses 3D printing approaches for fabricating
vascular networks and addresses the need for increased communication between
biologists and materials scientists.</div>
<div class="MsoNormal">
<br /></div>
<div class="MsoNormal">
WBC 2016 was an incredible conference in which I got to
present my work, learn about key advances in biomaterials, meet leaders in the
field, and explore Montreal. Thanks so much to RoosterBio for providing the
highest quality hMSCs and for their support of my work through a travel grant! <o:p></o:p></div>
Anonymousnoreply@blogger.com4tag:blogger.com,1999:blog-6181073719515632715.post-18494474477243112462016-06-14T15:05:00.002-04:002016-06-14T15:07:11.589-04:00World Biomaterials Congress 2016 Meeting Round Up Part I: Guest Blog Post<i>A guest blog post by RoosterBio Travel Award winner,<a href="http://millerlab.rice.edu/members/" target="_blank"> Gisele Calderon</a>.</i><br />
<i><br /></i>
<br />
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</v:shape><![endif]--><!--[if !vml]--><!--[endif]--><a href="https://3.bp.blogspot.com/-eghuYkEslvs/V2BUBBhmruI/AAAAAAAAD6s/N9yty4JN-DQsKbXzYbMlINn64PNF02LcgCLcB/s1600/Calderon1.jpg" imageanchor="1" style="clear: left; float: left; margin-bottom: 1em; margin-right: 1em;"><img border="0" height="320" src="https://3.bp.blogspot.com/-eghuYkEslvs/V2BUBBhmruI/AAAAAAAAD6s/N9yty4JN-DQsKbXzYbMlINn64PNF02LcgCLcB/s320/Calderon1.jpg" width="240" /></a><span style="font-family: "times new roman" , serif; font-size: 12.0pt;">The </span><a href="http://www.wbc2016.org/"><span style="font-family: "times new roman" , serif; font-size: 12.0pt;">World
Biomaterials Congress</span></a><span style="font-family: "times new roman" , serif; font-size: 12.0pt;"> (WBC) takes place every four years with an
energy rivaling the Olympics. This Congress is the largest gathering of biomaterials-focused
researchers with over </span><a href="http://tech4pco.com/wbc2016/mywbc/Abstract_Book/WBC2016_Abstracts.pdf"><span style="font-family: "times new roman" , serif; font-size: 12.0pt;">1,200
oral presentations and 2,400 poster presentations</span></a><span style="font-family: "times new roman" , serif; font-size: 12.0pt;">
representing over 60 countries. I am incredibly grateful to have been given the
opportunity to present my work to and learn from the World’s finest leaders of
the field. <o:p></o:p></span></div>
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<br /></div>
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</v:shape><![endif]--><!--[if !vml]--><!--[endif]--><span style="font-family: "times new roman" , serif; font-size: 12.0pt;">Our work
in the </span><a href="http://millerlab.rice.edu/"><span style="font-family: "times new roman" , serif; font-size: 12.0pt;">Miller Lab</span></a><span style="font-family: "times new roman" , serif; font-size: 12.0pt;"> at Rice
University focuses on vascularizing engineered tissues to address the metabolic
needs of these complex tissues via various techniques. I develop a system to
monitor cellular morphogenesis toward a stable capillary plexus allowing
biology to dictate the architectural hierarchy. The cell-cell interactions
between the endothelial cells derived from an iPS source and </span><a href="http://www.roosterbio.com/pages/what-are-mscs"><span style="font-family: "times new roman" , serif; font-size: 12.0pt;">human
mesenchymal stem cells</span></a><span style="font-family: "times new roman" , serif; font-size: 12.0pt;"> tend to enhance the stability of the putative
capillaries we form. Our novel multicolor genetic reporter system is enabling a
new class of longitudinal studies of tubulogenesis and their integration with
3D printed vasculature.<o:p></o:p></span></div>
<div class="MsoNormal" style="margin-bottom: 0.0001pt; text-align: justify;">
<br /></div>
<div class="separator" style="clear: both; text-align: center;">
<a href="https://3.bp.blogspot.com/-rbbgJvkDYUk/V2BUHfMqJPI/AAAAAAAAD60/OA7LsxjBzEIstv1F65r012h8QQOfRVfeACLcB/s1600/Calderon2.jpg" imageanchor="1" style="clear: right; float: right; margin-bottom: 1em; margin-left: 1em;"><img border="0" height="320" src="https://3.bp.blogspot.com/-rbbgJvkDYUk/V2BUHfMqJPI/AAAAAAAAD60/OA7LsxjBzEIstv1F65r012h8QQOfRVfeACLcB/s320/Calderon2.jpg" width="221" /></a></div>
<div class="MsoNormal" style="margin-bottom: 0.0001pt; text-align: justify;">
<span style="font-family: "times new roman" , serif; font-size: 12.0pt;">I was overjoyed to present my work to curious
peer grad students, esteemed thought-leaders, and industry representatives. I
was fortunately located near the coffee so my poster received a high amount of
traffic! I particularly enjoyed how accessible all of my science idols were
during the Congress. WBC ran an event where discussion and learning were the
central mission. <o:p></o:p></span></div>
<div class="MsoNormal" style="margin-bottom: 0.0001pt; text-align: justify;">
<br /></div>
<div class="MsoNormal" style="margin-bottom: 0.0001pt; text-align: justify;">
<span style="font-family: "times new roman" , serif; font-size: 12.0pt;">The Congress highlighted the field’s latest
work and how critical using therapeutically relevant cell types (like
RoosterBio’s hMSCs) is for successful tissue engineering strategies and forward
progress. I especially enjoyed engaging with investigators sharing the
following presentations:<o:p></o:p></span></div>
<div class="MsoNormal" style="margin-bottom: 0.0001pt; text-align: justify;">
<br /></div>
<div class="MsoNormal" style="margin-bottom: 0.0001pt; text-align: justify;">
<i style="text-indent: -0.25in;"><span style="font-family: "times new roman" , serif; font-size: 12.0pt;">3D
Tissue Printing</span></i></div>
<div class="MsoListParagraphCxSpFirst" style="margin-bottom: 0.0001pt; text-align: justify; text-indent: -0.25in;">
</div>
<ul>
<li><span style="font-family: "courier new"; font-size: 12pt; text-indent: -0.25in;"><span style="font-family: "times new roman"; font-size: 7pt; font-stretch: normal;"> </span></span><span style="font-family: "times new roman" , serif; font-size: 12pt; text-indent: -0.25in;">Dr.
Jennifer Lewis, </span><a href="http://lewisgroup.seas.harvard.edu/" style="text-indent: -0.25in;"><span style="font-family: "times new roman" , serif; font-size: 12.0pt;">Lewis
lab</span></a><span style="font-family: "times new roman" , serif; font-size: 12pt; text-indent: -0.25in;">, Wyss Institute, Harvard University</span></li>
<li><span style="font-family: "times new roman" , serif; font-size: 12pt; text-indent: -0.25in;"> How can
we directly print human tissue? Their approach utilizes top-down bioprinting
elegantly recreating complex vascular geometries.</span></li>
<li><span style="text-indent: -0.25in;"><span style="font-stretch: normal;"><span style="font-family: "times new roman" , serif;"> </span></span><span style="font-family: "times new roman"; font-size: 7pt; font-stretch: normal;"> </span></span><span style="font-family: "times new roman" , serif; font-size: 12pt; text-indent: -0.25in;">A recent
</span><a href="http://lewisgroup.seas.harvard.edu/files/lewisgroup/files/three-dimensional_bioprinting_of_thick_vascularized_tissues.pdf?m=1457464843" style="text-indent: -0.25in;"><span style="font-family: "times new roman" , serif; font-size: 12.0pt;">publication</span></a><span style="font-family: "times new roman" , serif; font-size: 12pt; text-indent: -0.25in;">
features RoosterBio’s hMSCs!</span></li>
</ul>
<i style="text-indent: -0.25in;"><span style="font-family: "times new roman" , serif; font-size: 12.0pt;"></span></i><br />
<div class="MsoListParagraphCxSpFirst" style="margin-bottom: 0.0001pt; text-align: justify; text-indent: -0.25in;">
<i style="text-indent: -0.25in;"><span style="font-family: "times new roman" , serif; font-size: 12.0pt;"><i style="text-indent: -0.25in;"><span style="font-family: "times new roman" , serif; font-size: 12.0pt;"><br /></span></i></span></i></div>
<i style="text-indent: -0.25in;"><span style="font-family: "times new roman" , serif; font-size: 12.0pt;">
3D Printing complex scaffolds using Freeform Reversible Embedding of Suspended
Hydrogels (FRESH)</span></i><br />
<div class="MsoListParagraphCxSpMiddle" style="margin-bottom: 0.0001pt; text-align: justify; text-indent: -0.25in;">
</div>
<ul>
<li><span style="font-family: "times new roman" , serif; font-size: 12pt; text-indent: -0.25in;"> Dr. Adam
Feinberg, </span><a href="http://regenerativebiomaterials.com/" style="text-indent: -0.25in;"><span style="font-family: "times new roman" , serif; font-size: 12.0pt;">Regenerative
Biomaterials & Therapeutics Group</span></a><span style="font-family: "times new roman" , serif; font-size: 12pt; text-indent: -0.25in;">, Carnegie Mellon University</span></li>
<li><span style="font-family: "times new roman" , serif; font-size: 12pt; text-indent: -0.25in;"> They 3DP
crazy complex structures in a gel within gel fashion for ubiquitous support in
their soft constructs. A fun note - he was inspired by Salvador Dali’s </span><a href="http://webneel.com/i/0/4-the-persistence-of-memory-surreal-art-by-salvador-dali/09-2013/d?n=9210" style="text-indent: -0.25in;"><span style="font-family: "times new roman" , serif; font-size: 12.0pt;">painting</span></a><span style="font-family: "times new roman" , serif; font-size: 12pt; text-indent: -0.25in;"> where
everything droops down without support!</span></li>
<li><span style="text-indent: -0.25in;"><span style="font-stretch: normal;"><span style="font-family: "times new roman" , serif;"> </span></span><span style="font-family: "times new roman"; font-size: 7pt; font-stretch: normal;"> </span></span><span style="font-family: "times new roman" , serif; font-size: 12pt; text-indent: -0.25in;">Here’s
their </span><a href="http://advances.sciencemag.org/content/advances/1/9/e1500758.full.pdf" style="text-indent: -0.25in;"><span style="font-family: "times new roman" , serif; font-size: 12.0pt;">FRESH
printing paper</span></a><span style="font-family: "times new roman" , serif; font-size: 12pt; text-indent: -0.25in;">.</span></li>
</ul>
<span style="font-family: "symbol"; font-size: 12pt; text-indent: -0.25in;"><span style="font-family: "times new roman"; font-size: 7pt; font-stretch: normal;"></span></span><br />
<div class="MsoListParagraphCxSpMiddle" style="margin-bottom: 0.0001pt; text-align: justify; text-indent: -0.25in;">
<span style="font-family: "symbol"; font-size: 12pt; text-indent: -0.25in;"><span style="font-family: "times new roman"; font-size: 7pt; font-stretch: normal;"><span style="font-family: "symbol"; font-size: 12pt; text-indent: -0.25in;"><span style="font-family: "times new roman"; font-size: 7pt; font-stretch: normal;"><br /></span></span></span></span></div>
<span style="font-family: "symbol"; font-size: 12pt; text-indent: -0.25in;"><span style="font-family: "times new roman"; font-size: 7pt; font-stretch: normal;">
</span></span><i style="text-indent: -0.25in;"><span style="font-family: "times new roman" , serif; font-size: 12.0pt;">Hydrogels
with continuously variable stiffness defined by dual-color
micro-stereolithography</span></i><br />
<div class="MsoListParagraphCxSpMiddle" style="margin-bottom: 0.0001pt; text-align: justify; text-indent: -0.25in;">
</div>
<ul>
<li><span style="font-family: "courier new"; font-size: 12pt; text-indent: -0.25in;"><span style="font-family: "times new roman"; font-size: 7pt; font-stretch: normal;"> </span></span><span style="font-family: "times new roman" , serif; font-size: 12pt; text-indent: -0.25in;">Dr.
Neils B Larsen, </span><a href="http://www.nanotech.dtu.dk/Research-mega/Forskningsgrupper/POLYCELL" style="text-indent: -0.25in;"><span style="font-family: "times new roman" , serif; font-size: 12.0pt;">PolyCell
group</span></a><span style="font-family: "times new roman" , serif; font-size: 12pt; text-indent: -0.25in;">, Technical University of Denmark</span></li>
<li><span style="font-family: "times new roman" , serif; font-size: 12pt; text-indent: -0.25in;"> This
group constructs 3DP soft constructs using stereolithography techniques in
order to incorporate microvessels in their hydrogels. They are able to achieve
consistent channels under 200um with tunable elasticity dependent on wavelength
and exposure tie of incident light.</span></li>
<li><span style="text-indent: -0.25in;"><span style="font-stretch: normal;"><span style="font-family: "times new roman" , serif;"> </span></span><span style="font-family: "times new roman"; font-size: 7pt; font-stretch: normal;"> </span></span><span style="font-family: "times new roman" , serif; font-size: 12pt; text-indent: -0.25in;">More
details can be found </span><a href="http://onlinelibrary.wiley.com/doi/10.1002/polb.24007/full" style="text-indent: -0.25in;"><span style="font-family: "times new roman" , serif; font-size: 12.0pt;">here</span></a><span style="font-family: "times new roman" , serif; font-size: 12pt; text-indent: -0.25in;">.</span></li>
</ul>
<br />
<br />
<div class="MsoNormal" style="margin-bottom: 0.0001pt; text-align: justify;">
<span style="font-family: "times new roman" , serif; font-size: 12.0pt;">In reality, there are details of many more presentations
that I would love to share here, but I just couldn’t do justice to the high
spirit of scientific rigor. Throughout the Congress, I was actively tweeting
about all the excellent work. Follow me </span><a href="https://twitter.com/g_caldero"><span style="font-family: "times new roman" , serif; font-size: 12.0pt;">@g_caldero</span></a><span style="font-family: "times new roman" , serif; font-size: 12.0pt;">! And
lastly, thank you, RoosterBio, for the awesome cells AND also the travel award
support to attend this exciting Congress! <o:p></o:p></span></div>
Anonymousnoreply@blogger.com0tag:blogger.com,1999:blog-6181073719515632715.post-2793024785104399092016-05-10T07:52:00.000-04:002016-05-10T07:52:21.456-04:00Comparability of hMSC Economic and Quality Attributes after Expansion in Bovine Serum Containing vs Xeno-Free Bioprocessing Media Formulations <div class="MsoNormal">
<a href="http://www.roosterbio.com/pages/what-are-mscs">Human
Mesenchymal Stem/Stromal Cells (hMSCs)</a>, from bone marrow or other tissues, are
poised to have the most significant impact on Regenerative Medicine compared to
any other <u>single</u> cell type. This
is due to their ability to be utilized across multiple therapeutic indications
due to the wide ranging functional nature of the cells (1-3). hMSCs are not only capable of differentiating
into tissue-specific cell types, but also have angiogenic, immunomodulatory,
anti-inflammatory and anti-bacterial abilities (4). <a href="http://roosterbio.blogspot.com/2014/03/mesenchymal-stem-cells-workhorse-of.html">hMSCs
are true Tissue Repair Cells</a> – setting the stage for all phases of wound
healing and tissue repair: promoting new blood vessel growth, reducing
inflammation to aid healing, secreting several mitogenic factors important for
tissue building and stimulating tissue-specific stem cells. <o:p></o:p></div>
<div class="MsoNormal">
<br /></div>
<div class="MsoNormal">
However, hMSCs have traditionally been challenging to source
in significant volumes and at sufficient quality levels, <a href="http://roosterbio.blogspot.com/2014/02/current-bottlenecks-in-msc-research.html">hindering
the advancement of the science</a> into medical products. At RoosterBio, we focus on transitioning
hMSCs from a scarce into an abundant resource, and we achieve this by borrowing
best practices from the Manufacturing Sciences and applying them towards the
grand challenge of producing billions of hMSCs, with critical quality and
functional parameters in place, and at costs and volumes that enable the rapid
and wide-spread adoption of hMSC technology into clinical practice.<o:p></o:p></div>
<div class="MsoNormal">
<br /></div>
<div class="MsoNormal">
RoosterBio came to market 2 years ago with hMSC cell and
media systems that include a highly efficient <a href="http://www.roosterbio.com/collections/media/products/hbm-msc-high-productivity-media-kit">hMSC
bioprocess expansion media</a> that
simply and consistently produces greater than 100x expansion of cells with 8-10
days of culture. Our cell and media system was designed for a “batch” culture
process (no media exchange required between passages), removing labor-intensive
and costly media exchanges, and enabling rapid expansion with little in process
intervention (thus fewer risks for contamination). While the cell and media system has now been
used in several translational and high impact publications (5-8), the expansion
medium does utilize low levels of high quality bovine serum to maximize the performance
and robustness of the overall system. <o:p></o:p></div>
<div class="MsoNormal">
<br /></div>
<div class="MsoNormal">
In recent years, the field has been shifting towards xeno-free
(XF) cell and media systems to remove any remaining safety issues related to xeno-sourced
animal components (9-13). Furthermore, our customers have been requesting XF
expansion options. We have listened to our customers and spent the last year
developing and optimizing a fully XF media formulation based on our innovative bioprocess
media platform. The goals of this media
were to remove all xeno-sourced raw materials from the formulation, while
maintaining all hMSC functional properties, as well as the economic and production
efficiency of our initial bovine serum containing (BSC) media formulation. We are now ready to commercially launch our XF
media to advance the industry, and this blog post will outline the initial work
we have performed to evaluate the comparability of expansion, cost and
functional properties of hMSCs expanded in the new XF media compared to our flagship
BSC media.<o:p></o:p></div>
<div class="separator" style="clear: both; text-align: center;">
</div>
<table align="center" cellpadding="0" cellspacing="0" class="tr-caption-container" style="margin-left: auto; margin-right: auto; text-align: center;"><tbody>
<tr><td style="text-align: center;"><a href="https://1.bp.blogspot.com/-sWwozq6V5KQ/VzHKaw_NqOI/AAAAAAAAD4g/n-zvaGRXVaMWwys5A_In47lyhbdA5HnRQCLcB/s1600/Table%2B1.png" imageanchor="1" style="margin-left: auto; margin-right: auto;"><img border="0" height="166" src="https://1.bp.blogspot.com/-sWwozq6V5KQ/VzHKaw_NqOI/AAAAAAAAD4g/n-zvaGRXVaMWwys5A_In47lyhbdA5HnRQCLcB/s640/Table%2B1.png" width="640" /></a></td></tr>
<tr><td class="tr-caption" style="text-align: center;"><span style="font-family: "times" , "times new roman" , serif; font-size: small;"><b>Table 1.</b> <i>Media formulations and nomenclature.</i></span></td></tr>
</tbody></table>
<div class="separator" style="clear: both; text-align: center;">
</div>
For the purpose of this blog post, we will be comparing
RoosterBio hMSC products expanded in either our initial bovine serum containing
<a href="http://www.roosterbio.com/collections/media/products/hbm-msc-high-productivity-media-kit">High
Performance Media</a>, or our
new xeno-free <a href="http://www.roosterbio.com/collections/media/products/hmsc-high-performance-media-kit-xf-kt-016" target="_blank">High Performance Media XF</a> formulation.<br />
<div class="MsoNormal">
<o:p></o:p></div>
<div class="MsoNormal">
<br /></div>
<div class="MsoNormal">
<b><span style="color: #1f497d; mso-themecolor: text2;"><span style="color: blue;">METHODS</span><span style="color: #1f497d;"><o:p></o:p></span></span></b></div>
<div class="MsoNormal">
<br /></div>
<div class="MsoNormal">
<b><i>Cell expansion.</i></b><i> </i>RoosterBio
hBM-MSC were expanded in BSC Media and XF Media. Frozen cells were thawed and
plated in triplicate at 3,000 cells/cm<sup>2</sup> in T-75 flasks and cultured
for 4 days. At 4 days, cells were harvested with TrypLE (Gibco) and cell number
and viability were determined on a Nucleocounter. These cells were used for the
analyses below or plated again for further expansion.</div>
<div class="MsoNormal">
<o:p></o:p></div>
<div class="MsoNormal">
<b><i><br /></i></b></div>
<div class="MsoNormal">
<b><i>Cell surface marker expression.</i></b> To determine if the cells grown
in XF Media were capable of expressing MSC markers, hBM-MSC expanded in both
BSC and XF Media were plated and incubated in DMEM/10% FBS for 5 days prior to
flow cytometry. <o:p></o:p></div>
<div class="MsoNormal">
<b><i><br /></i></b></div>
<div class="MsoNormal">
<b><i>Immunomodulatory function</i></b><i>.</i>
Induction of indoleamine 2,3-dioxygenase (IDO) activity by exposure of hMSCs to
the pro-inflamatory cytokine IFN-γ is central to the immunosuppressive function
of hMSCs (14,15). See <a href="http://roosterbio.blogspot.com/2014/12/priming-of-hmscs-to-improve-potency.html">here</a>
for a blog post on this topic. hBM-MSCs were expanded in BSC and XF Media
(Donors 1 and 2) or XF Media alone (Donor 3), harvested and plated in High
Performance Basal medium (SU-005) with 2% FBS at 40,000 cells/cm<sup>2</sup>.
After 18-22 hr of incubation, cells were treated with IFN-γ (10 ng/ml) for
24hr±1hr. The cell supernatant was collected, and the kynurenine concentration
was measured using a spectrophotometric assay and normalized to number of cells
and days of incubation to obtain the amount of IDO secreted (expressed as pg
kynurenine secreted per cell per day). <o:p></o:p></div>
<div class="MsoNormal">
<b><i><br /></i></b></div>
<div class="MsoNormal">
<b><i>Angiogenic cytokine secretion.</i></b><i> </i>hBM-MSCs were expanded in BSC or XF Media, harvested and plated in
High Performance Basal Medium with 2% FBS at 40,000 cells/cm<sup>2</sup>. After
24hr±1hr culture supernatant was collected and assayed for FGF, HGF, IL-8,
TIMP-1, TIMP-2 and VEGF concentration using a MultiPlex ELISA (Quansys).
Cytokine concentration was normalized to number of cells and days of incubation
to obtain cytokine secretion rates.<o:p></o:p></div>
<div class="MsoNormal">
<b><i><br /></i></b></div>
<div class="MsoNormal">
<b><i>Trilineage differentiation.</i></b> hBM-MSCs were expanded in BSC or XF
Media, harvested and plated in High Performance Basal Medium with 2% FBS at 5,000-10,000
cells/cm<sup>2 </sup> for adipogenesis
and osteogenesis or formed into 100,000 cell micromasses for chondrogenesis. On
day 1, cells were switched to differentiation or control media (LifeTech
StemPro Differentiation Kits) and cultured per kit protocols for 10-21 days. Differentiation
was detected by Oil Red O (adipogenesis), Alizarin Red (osteogenesis), or
Toluidine Blue (chondrogenesis) stains. <o:p></o:p></div>
<div class="MsoNormal">
<span style="color: blue;"><br /></span></div>
<div class="MsoNormal">
<b><span style="color: #1f497d; mso-themecolor: text2;"><span style="color: blue;">COMPARATIVE ANALYSES</span><span style="color: #1f497d;"><o:p></o:p></span></span></b></div>
<div class="MsoNormal">
<b><i><br /></i></b></div>
<div class="MsoNormal">
<b><i>Cell expansion.</i></b><i> </i>A key
characteristic of RoosterBio hMSC cell and media systems is rapid cell expansion
with a guaranteed 10-fold expansion within 7 days. In engineering our XF Media
system, we aimed to preserve this hMSC expansion profile. hBM-MSCs display rapid and comparable growth
in both our BSC Media and the new XF Media formulations, with similar doubling
times and expansion rates. We see the
typical variability across donors, but all donors are harvested at greater than
30,000 cells/cm<sup>2</sup>, after plating at 3,000 cells/cm<sup>2</sup>,
within 5 days (Figure 1). hBM-MSC growth over 2 passages yields greater than 1 billion
cells using both our BSC and XF Media (and <a href="http://www.roosterbio.com/collections/cells-1">10M cell product vials</a>)
in less than 2 weeks (Figure 2), leading to tremendous economic benefits
(described below).<br />
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<tr><td style="text-align: center;"><a href="https://1.bp.blogspot.com/-kyewDt-LMHQ/VzE8UuSOGkI/AAAAAAAAD20/gVLLjK6hrmI8yGWrPCflPKwwziUmVlKHgCLcB/s1600/Fig1.jpg" imageanchor="1" style="clear: right; margin-bottom: 1em; margin-left: auto; margin-right: auto;"><img border="0" height="291" src="https://1.bp.blogspot.com/-kyewDt-LMHQ/VzE8UuSOGkI/AAAAAAAAD20/gVLLjK6hrmI8yGWrPCflPKwwziUmVlKHgCLcB/s320/Fig1.jpg" width="320" /></a></td></tr>
<tr><td class="tr-caption" style="text-align: center;"><div style="text-align: left;">
<span style="font-family: "times" , "times new roman" , serif; font-size: small;"><b>Figure 1.</b> <span style="font-style: italic; text-align: left;">hBM</span><span style="font-style: italic; text-align: left;">-MSCs expand efficiently in XF Media</span><span style="text-align: left;">.<i> </i></span><span style="text-align: left;">All</span><span style="text-align: left; vertical-align: baseline;"> </span><span style="text-align: left;">cell lots expanded at least 10-fold
(>30,000</span><span style="text-align: left; vertical-align: baseline;"> cells/cm</span><span style="text-align: left; vertical-align: super;">2</span><span style="text-align: left; vertical-align: baseline;">) </span><span style="text-align: left;">in XF</span><span style="text-align: left; vertical-align: baseline;"> and BSC Media (data</span><span style="text-align: left; vertical-align: baseline;"> shown for 2 donor lots). </span><span style="text-align: left;">Data
are mean of</span><span style="text-align: left; vertical-align: baseline;"> 3 replicates +/- SD.</span></span><br />
<a name='more'></a></div>
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<tr><td style="text-align: center;"><a href="https://1.bp.blogspot.com/-AWdYSz-jz2k/VzE9pMI6hbI/AAAAAAAAD3A/k5L5NXjcQOwVSVkZkBZ_sGx6VLE_-EnoACLcB/s1600/Fig2.jpg" imageanchor="1" style="clear: right; margin-bottom: 1em; margin-left: auto; margin-right: auto;"><img border="0" height="227" src="https://1.bp.blogspot.com/-AWdYSz-jz2k/VzE9pMI6hbI/AAAAAAAAD3A/k5L5NXjcQOwVSVkZkBZ_sGx6VLE_-EnoACLcB/s320/Fig2.jpg" width="320" /></a></td></tr>
<tr><td class="tr-caption" style="text-align: left;"><span style="font-family: "times" , "times new roman" , serif; font-size: small;"><b>Figure 2. </b><span style="font-style: italic; text-align: left; vertical-align: baseline;">RoosterBio</span><span style="font-style: italic; text-align: left; vertical-align: baseline;"> BSC </span><span style="font-style: italic; text-align: left; vertical-align: baseline;">Media and XF Media quickly generate
>100B hMSC</span><span style="font-style: italic; text-align: left;">. </span><span style="text-align: left;">RoosterBio</span><span style="text-align: left;"> </span><span style="text-align: left;">cells in both BSC and XF Media showed a
200-fold expansion in 8 days vs. 10-</span><span style="text-align: left; vertical-align: baseline;"> to
20</span><span style="text-align: left;">-fold
expansion rate in</span><span style="text-align: left; vertical-align: baseline;"> 14 </span><span style="text-align: left; vertical-align: baseline;">days
(over 2 passages) in</span><span style="text-align: left; vertical-align: baseline;"> other Suppliers' systems.</span></span></td></tr>
</tbody></table>
<b><i>Cell surface marker expression. </i></b>Flow cytometry analyses of hBM-MSC
cell surface marker expression in XF and BSC Media demonstrated no differences
between the two cell populations. hBM-MSC populations cultured in both media
formulations were low for hematopoietic stem cell markers, CD14, CD45 and CD34,
and >90% positive for hMSC markers CD166, CD105, CD90, and CD73 (Figure 3). <o:p></o:p></div>
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<tr><td class="tr-caption" style="text-align: left;"><span style="font-family: "times" , "times new roman" , serif; font-size: small;"><b>Figure 3. </b><span style="font-style: italic; text-align: left;">XF</span><span style="font-style: italic; text-align: left; vertical-align: baseline;">
Media</span><span style="font-style: italic; text-align: left;">
does not change </span><span style="font-style: italic; text-align: left;">hMSC</span><span style="font-style: italic; text-align: left;"> cell surface marker expression.</span><span style="text-align: left;">
Cells from both media types were >95% positive for CD166, CD105, CD90 and
CD73 and <5% positive for CD14, CD34 and CD45. Identical</span><span style="text-align: left; vertical-align: baseline;"> results were obtained with other Donors
1 and 3 (data not shown).</span></span></td></tr>
</tbody></table>
<b><i>Immunomodulatory function. </i></b>While significant donor-to-donor
variation in IDO induction levels post-IFN-γ treatment exists in hMSCs, hBM-MSCs
that were expanded in both BSC and XF Media in parallel showed similar IDO
activity after IFN-γ treatment (Figure 4). All cell/donor lots that were
expanded in XF Media showed inducible IDO activity (some donors shown in Figure
4) thereby confirming that cells maintain critical hMSC immunomodulatory
function in both media formulations.<o:p></o:p></div>
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<tr><td style="text-align: center;"><a href="https://4.bp.blogspot.com/-9aQjDqdgzVQ/VzE-6dhUFKI/AAAAAAAAD3Q/fcpozH46a7wEc7rhMb1MV28iThPKLy-8gCLcB/s1600/Fig4.jpg" imageanchor="1" style="margin-left: auto; margin-right: auto;"><img border="0" height="238" src="https://4.bp.blogspot.com/-9aQjDqdgzVQ/VzE-6dhUFKI/AAAAAAAAD3Q/fcpozH46a7wEc7rhMb1MV28iThPKLy-8gCLcB/s320/Fig4.jpg" width="320" /></a></td></tr>
<tr><td class="tr-caption" style="text-align: center;"><div style="text-align: left;">
<span style="font-size: small;"><b style="font-family: times, 'times new roman', serif;">Figure 4. </b><span style="font-family: "times" , "times new roman" , serif; font-style: italic; text-align: left; text-indent: 0in;">I</span><span style="font-family: "times" , "times new roman" , serif; font-style: italic; text-align: left; text-indent: 0in;">mmunomodulatory</span><span style="font-family: "times" , "times new roman" , serif; font-style: italic; text-align: left; text-indent: 0in; vertical-align: baseline;"> activity </span><span style="font-family: "times" , "times new roman" , serif; font-style: italic; text-align: left; text-indent: 0in; vertical-align: baseline;">of </span><span style="font-family: "times" , "times new roman" , serif; font-style: italic; text-align: left; text-indent: 0in; vertical-align: baseline;">hMSCs</span><span style="font-family: "times" , "times new roman" , serif; font-style: italic; text-align: left; text-indent: 0in; vertical-align: baseline;"> is </span><span style="font-family: "times" , "times new roman" , serif; font-style: italic; text-align: left; text-indent: 0in; vertical-align: baseline;">similar in </span><span style="font-family: "times" , "times new roman" , serif; font-style: italic; text-align: left; text-indent: 0in; vertical-align: baseline;">BSC </span><span style="font-family: "times" , "times new roman" , serif; font-style: italic; text-align: left; text-indent: 0in; vertical-align: baseline;">and XF Media</span><span style="font-family: "times" , "times new roman" , serif; font-style: italic; text-align: left; text-indent: 0in;">.</span><span style="font-family: "times" , "times new roman" , serif; text-align: left; text-indent: 0in;"> </span><span style="font-family: "times" , "times new roman" , serif; text-align: left; text-indent: 0in; vertical-align: baseline;">Donors that were expanded in both BSC and
XF Media in parallel showed similar IDO activity after I</span><span style="font-family: "times" , "times new roman" , serif; text-align: left; text-indent: 0in;">FN-</span><span style="font-family: "times" , "times new roman" , serif; text-align: left; text-indent: 0in;">γ</span><span style="font-family: "times" , "times new roman" , serif; text-align: left; text-indent: 0in;">
treatment</span><span style="font-family: "times" , "times new roman" , serif; text-align: left; text-indent: 0in; vertical-align: baseline;">. A</span><span style="font-family: "times" , "times new roman" , serif; text-align: left; text-indent: 0in;">ll</span><span style="font-family: "times" , "times new roman" , serif; text-align: left; text-indent: 0in; vertical-align: baseline;">
Donors</span><span style="font-family: "times" , "times new roman" , serif; text-align: left; text-indent: 0in;"> that
were</span><span style="font-family: "times" , "times new roman" , serif; text-align: left; text-indent: 0in; vertical-align: baseline;"> </span><span style="font-family: "times" , "times new roman" , serif; text-align: left; text-indent: 0in;">expanded in XF Media showed inducible</span><span style="font-family: "times" , "times new roman" , serif; text-align: left; text-indent: 0in; vertical-align: baseline;"> IDO activity </span><span style="font-family: "times" , "times new roman" , serif; text-align: left; text-indent: 0in; vertical-align: baseline;">(not </span><span style="font-family: "times" , "times new roman" , serif; text-align: left; text-indent: 0in; vertical-align: baseline;">shown).
</span><span style="font-family: "times" , "times new roman" , serif; text-align: left; text-indent: 0in;">The</span><span style="font-family: "times" , "times new roman" , serif; text-align: left; text-indent: 0in; vertical-align: baseline;"> IDO activity for untreated cells was
from 0 to 0.12 </span><span style="font-family: "times" , "times new roman" , serif; text-align: left; text-indent: 0in; vertical-align: baseline;">pg</span><span style="font-family: "times" , "times new roman" , serif; text-align: left; text-indent: 0in; vertical-align: baseline;"> </span><span style="font-family: "times" , "times new roman" , serif; text-align: left; text-indent: 0in; vertical-align: baseline;">kynurenine</span><span style="font-family: "times" , "times new roman" , serif; text-align: left; text-indent: 0in; vertical-align: baseline;">/cell/day
(not shown).</span></span></div>
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<b><i>Angiogenic cytokine secretion
profile. </i></b>hMSC achieve their therapeutic effects primarily by secreting
a plethora of biomolecules that influence many biologic processes (2,3). We
assay the secretion of bFGF, HGF, TIMP 1, TIMP 2, IL-8, and VEGF via
multiplexed ELISA analyses as part of
our standard QC assays for each hMSC lot we manufacture. We assayed cytokine
secretion of 2 donors grown in XF Media, two of which were also assayed after
expansion in BSC Media (Figure 5). Comparable cytokine secretion profiles were
observed for cells grown with XF and BSC Media, all within donor-to-donor
variation. <o:p></o:p></div>
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<tr><td style="text-align: center;"><a href="https://1.bp.blogspot.com/-KjCDwRpj4mw/VzE_xxOa9aI/AAAAAAAAD3g/0N0NlSYSJcMhyWYh82r_7ljPTJvaIwf3QCLcB/s1600/Fig5.jpg" imageanchor="1" style="margin-left: auto; margin-right: auto;"><img border="0" height="226" src="https://1.bp.blogspot.com/-KjCDwRpj4mw/VzE_xxOa9aI/AAAAAAAAD3g/0N0NlSYSJcMhyWYh82r_7ljPTJvaIwf3QCLcB/s400/Fig5.jpg" width="400" /></a></td></tr>
<tr><td class="tr-caption" style="text-align: center;"><div style="text-align: left;">
<span style="font-family: "times" , "times new roman" , serif; font-size: small;"><b>Figure 5. </b><span style="font-style: italic; text-align: left;">Angiogenic</span><span style="font-style: italic; text-align: left; vertical-align: baseline;"> c</span><span style="font-style: italic; text-align: left;">ytokine secretion is maintained.</span><span style="text-align: left;">
Culture supernatant from </span><span style="text-align: left;">hBM</span><span style="text-align: left;">-MSC
(2</span><span style="text-align: left; vertical-align: baseline;"> Donors) </span><span style="text-align: left;">grown</span><span style="text-align: left; vertical-align: baseline;"> in
BSC and XF Media </span><span style="text-align: left;">demonstrated comparable secretion of FGF, HGF, IL-8, TIMP-1, TIMP-2 and VEGF.</span></span></div>
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<b><i>Trilineage differentiation</i></b><i>.
</i>A hallmark characteristic of hMSCs is <i>in vitro</i> differentiation to osteoblasts, adipocytes and
chondrocytes (16,17). hBM-MSCs grown in XF Media differentiated to fat, bone
and cartilage, with no qualitative differences compared to cells grown in BSC Media
(Figure 6).<o:p></o:p></div>
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<tr><td style="text-align: center;"><a href="https://3.bp.blogspot.com/-sOFj9sBxf_w/VzFAKmO5RvI/AAAAAAAAD3k/Q5UoUkvaXh03BF8vlVkTieTHKq4QbtelACLcB/s1600/Fig6.jpg" imageanchor="1" style="margin-left: auto; margin-right: auto;"><img border="0" height="356" src="https://3.bp.blogspot.com/-sOFj9sBxf_w/VzFAKmO5RvI/AAAAAAAAD3k/Q5UoUkvaXh03BF8vlVkTieTHKq4QbtelACLcB/s640/Fig6.jpg" width="640" /></a></td></tr>
<tr><td class="tr-caption" style="text-align: left;"><span style="font-size: small;"><span style="font-family: "times" , "times new roman" , serif; font-size: xx-small;"><b>Figure 6. </b><i><span style="text-align: left; text-indent: 0in; vertical-align: baseline;">hBM-MSCs</span><span style="text-align: left; text-indent: 0in; vertical-align: baseline;"> were
capable of </span><span style="text-align: left; text-indent: 0in;">t</span><span style="text-align: left; text-indent: 0in;">rilineage</span><span style="text-align: left; text-indent: 0in; vertical-align: baseline;"> d</span><span style="text-align: left; text-indent: 0in;">ifferentiation
(</span><span style="text-align: left; text-indent: 0in;">adipo</span><span style="text-align: left; text-indent: 0in;">-,</span><span style="text-align: left; text-indent: 0in; vertical-align: baseline;"> </span><span style="text-align: left; text-indent: 0in; vertical-align: baseline;">osteo</span><span style="text-align: left; text-indent: 0in; vertical-align: baseline;">- </span><span style="text-align: left; text-indent: 0in;">and </span><span style="text-align: left; text-indent: 0in;">chondrogenesis</span><span style="text-align: left; text-indent: 0in;">) </span></i></span><i style="font-family: times, 'times new roman', serif;">in both BSC and XF Media </i><i style="font-family: times, 'times new roman', serif;"><span style="text-indent: 0in;">.</span></i></span></td></tr>
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<b>In summary, the
critical quality attributes of RoosterBio hBM-MSCs are maintained and are comparable
between cells expanded in RoosterBio’s High Performance Media and our new XF
Media formulation. <o:p></o:p></b></div>
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<b><span style="color: #1f497d; mso-themecolor: text2;"><span style="color: blue;">COST AND YIELD ANALYSES</span><span style="color: #1f497d;"><o:p></o:p></span></span></b></div>
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The time and economic savings that both RoosterBio BSC and XF
Media formulations afford are the major factor in customers’ switching to RoosterBio
hMSC culture systems. Using the yield
and labor data from Figure 2 enables the demonstration of significant cost and
time savings for generating 100 Million cells for applications or experiments.
These innovative RoosterBio cell and media systems help turn scientific experiments
into commercially feasible technologies for implementation.<o:p></o:p></div>
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<tr><td style="text-align: center;"><a href="https://4.bp.blogspot.com/-jBT4DSM3baA/VzFBHUuIwMI/AAAAAAAAD34/hq0an1oSDhQSY6Q9lrYJ4ZUTIzQFKoU4QCLcB/s1600/Table%2B2.png" imageanchor="1" style="margin-left: auto; margin-right: auto;"><img border="0" height="287" src="https://4.bp.blogspot.com/-jBT4DSM3baA/VzFBHUuIwMI/AAAAAAAAD34/hq0an1oSDhQSY6Q9lrYJ4ZUTIzQFKoU4QCLcB/s640/Table%2B2.png" width="640" /></a></td></tr>
<tr><td class="tr-caption" style="text-align: left;"><span style="font-family: "times" , "times new roman" , serif; font-size: small;"><b>Table 2. </b><i>Cost and time analyses</i> for generating 100 Million viable
cells from the leading hMSC and media systems available commercially today.
Time to cells in days.</span></td></tr>
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The most significant aspect of the cost analysis is the
savings in media volume. Media has
always been a major cost driver in biomanufacturing processes. Just as bioprocess innovations
have enhanced the productivity in monoclonal antibody production and driven
exponential cost savings in biotherapeutics, we are driving similar
efficiencies with our bioprocess media systems.
The key is media productivity, and when the metric of “Millions of Cells
produced per Liter of Media” is evaluated, the differences are illustrative of
massive manufacturing enhancements. Figure 7 shows that the there is
essentially a 9x enhancement in million cells produced per liter of media
utilized in culture expansion with RoosterBio media systems versus others. <o:p></o:p></div>
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<tr><td style="text-align: center;"><a href="https://4.bp.blogspot.com/-r03UyhMuZWg/VzFB6iB_wQI/AAAAAAAAD4I/HBrcybvrN1Q0vq7TWNJv1TsOWUOBvokwACLcB/s1600/Fig7.jpg" imageanchor="1" style="margin-left: auto; margin-right: auto;"><img border="0" height="265" src="https://4.bp.blogspot.com/-r03UyhMuZWg/VzFB6iB_wQI/AAAAAAAAD4I/HBrcybvrN1Q0vq7TWNJv1TsOWUOBvokwACLcB/s400/Fig7.jpg" width="400" /></a></td></tr>
<tr><td class="tr-caption" style="text-align: center;"><div style="text-align: left;">
<span style="font-family: "times" , "times new roman" , serif; font-size: small;"><b>Figure 7. </b><i>RoosterBio systems yield 9x more cells per Liter of media consumed than other cell and media systems.</i></span></div>
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Of course, Quality is always the most critical metric, and
if cell functionality is compromised, then any cost enhancement cannot be
justified. However, the functional data
we have generated clearly shows that hMSC functions are maintained in our
systems. Our aim in commercializing
these systems is to enable the next generation of medical devices that include
human stem cells to rapidly progress through product development and clinical
development.<o:p></o:p></div>
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We believe we are seeing the dawn of a new era of productivity
and product development in the Regenerative Medicine field. If there are any other innovations you think
will have a dramatic impact on the field, please comment below.<o:p></o:p></div>
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<b><span style="font-size: x-small;">REFERENCES<o:p></o:p></span></b></div>
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SH, Levy O, Inamdar MS, & Karp JM (2012) Harnessing the mesenchymal stem
cell secretome for the treatment of cardiovascular disease. <i>Cell stem cell</i> 10(3):244-258. </a><a href="http://www.ncbi.nlm.nih.gov/pubmed/22385653">http://www.ncbi.nlm.nih.gov/pubmed/22385653</a><o:p></o:p></span></div>
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AM, Caplan AI, & Bonfield TL (2013) Mesenchymal stem cells in tissue
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DB, Homan KA, Skylar-Scott MA, & Lewis JA (2016) Three-dimensional
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O, Brennen WN, Han E, Rosen DM, Musabeyezu J, Safaee H, Ranganath S, Ngai J,
Heinelt M, Milton Y, Wang H, Bhagchandani SH, Joshi N, Bhowmick N, Denmeade SR,
Isaacs JT, & Karp JM (2016) A prodrug-doped cellular Trojan Horse for the
potential treatment of prostate cancer. <i>Biomaterials</i>
91:140-150. </a><a href="http://www.ncbi.nlm.nih.gov/pubmed/27019026">http://www.ncbi.nlm.nih.gov/pubmed/27019026</a><o:p></o:p></span></div>
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<span style="font-size: x-small;"><a href="https://www.blogger.com/null" name="_ENREF_7">7. Nguyen
BN, Ko H, Moriarty RA, Etheridge JM, & Fisher JP (2016) Dynamic Bioreactor
Culture of High Volume Engineered Bone Tissue. <i>Tissue engineering. Part A</i> 22(3-4):263-271. </a><a href="http://www.ncbi.nlm.nih.gov/pubmed/26653703">http://www.ncbi.nlm.nih.gov/pubmed/26653703</a><o:p></o:p></span></div>
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<span style="font-size: x-small;"><a href="https://www.blogger.com/null" name="_ENREF_8">8. Tan
KY, Teo KL, Lim JF, Chen AK, Choolani M, Reuveny S, Chan J, & Oh SK (2015)
Serum-free media formulations are cell line-specific and require optimization
for microcarrier culture. <i>Cytotherapy</i>
17(8):1152-1165. </a><a href="http://www.ncbi.nlm.nih.gov/pubmed/26139547">http://www.ncbi.nlm.nih.gov/pubmed/26139547</a><o:p></o:p></span></div>
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<span style="font-size: x-small;"><a href="https://www.blogger.com/null" name="_ENREF_9">9. Copland
IB, Garcia MA, Waller EK, Roback JD, & Galipeau J (2013) The effect of
platelet lysate fibrinogen on the functionality of MSCs in immunotherapy. <i>Biomaterials</i> 34(32):7840-7850. </a><a href="http://www.ncbi.nlm.nih.gov/pubmed/23891515">http://www.ncbi.nlm.nih.gov/pubmed/23891515</a><o:p></o:p></span></div>
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<span style="font-size: x-small;"><a href="https://www.blogger.com/null" name="_ENREF_10">10. Escobar CH & Chaparro O (2016) Xeno-Free Extraction,
Culture, and Cryopreservation of Human Adipose-Derived Mesenchymal Stem Cells. <i>Stem cells translational medicine</i>
5(3):358-365. </a><a href="http://www.ncbi.nlm.nih.gov/pubmed/26838269">http://www.ncbi.nlm.nih.gov/pubmed/26838269</a><o:p></o:p></span></div>
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<span style="font-size: x-small;"><a href="https://www.blogger.com/null" name="_ENREF_11">11. Heathman TR, Stolzing A, Fabian C, Rafiq QA, Coopman K, Nienow
AW, Kara B, & Hewitt CJ (2015) Serum-free process development: improving
the yield and consistency of human mesenchymal stromal cell production. <i>Cytotherapy</i> 17(11):1524-1535. </a><a href="http://www.ncbi.nlm.nih.gov/pubmed/26432558">http://www.ncbi.nlm.nih.gov/pubmed/26432558</a><o:p></o:p></span></div>
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<span style="font-size: x-small;"><a href="https://www.blogger.com/null" name="_ENREF_12">12. Heathman TR, Stolzing A, Fabian C, Rafiq QA, Coopman K, Nienow
AW, Kara B, & Hewitt CJ (2016) Scalability and process transfer of
mesenchymal stromal cell production from monolayer to microcarrier culture
using human platelet lysate. <i>Cytotherapy</i>
18(4):523-535. </a><a href="http://www.ncbi.nlm.nih.gov/pubmed/26971681">http://www.ncbi.nlm.nih.gov/pubmed/26971681</a><o:p></o:p></span></div>
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<span style="font-size: x-small;"><a href="https://www.blogger.com/null" name="_ENREF_13">13. Oikonomopoulos A, van Deen WK, Manansala AR, Lacey PN,
Tomakili TA, Ziman A, & Hommes DW (2015) Optimization of human mesenchymal
stem cell manufacturing: the effects of animal/xeno-free media. <i>Sci Rep</i> 5:16570. </a><a href="http://www.ncbi.nlm.nih.gov/pubmed/26564250">http://www.ncbi.nlm.nih.gov/pubmed/26564250</a><o:p></o:p></span></div>
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<span style="font-size: x-small;"><a href="https://www.blogger.com/null" name="_ENREF_14">14. Francois M & Galipeau J (2012) New insights on
translational development of mesenchymal stromal cells for suppressor therapy. <i>Journal of cellular physiology</i>
227(11):3535-3538. </a><a href="http://www.ncbi.nlm.nih.gov/pubmed/22378308">http://www.ncbi.nlm.nih.gov/pubmed/22378308</a><o:p></o:p></span></div>
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<span style="font-size: x-small;"><a href="https://www.blogger.com/null" name="_ENREF_15">15. Krampera M, Galipeau J, Shi Y, Tarte K, & Sensebe L (2013)
Immunological characterization of multipotent mesenchymal stromal cells--The
International Society for Cellular Therapy (ISCT) working proposal. <i>Cytotherapy</i> 15(9):1054-1061. </a><a href="http://www.ncbi.nlm.nih.gov/pubmed/23602578">http://www.ncbi.nlm.nih.gov/pubmed/23602578</a><o:p></o:p></span></div>
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<span style="font-size: x-small;"><a href="https://www.blogger.com/null" name="_ENREF_16">16. Pittenger MF (2008) Mesenchymal stem cells from adult bone
marrow. <i>Methods in molecular biology</i>
449:27-44. </a><a href="http://www.ncbi.nlm.nih.gov/pubmed/18370081">http://www.ncbi.nlm.nih.gov/pubmed/18370081</a><o:p></o:p></span></div>
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<span style="font-size: x-small;"><a href="https://www.blogger.com/null" name="_ENREF_17"><span style="mso-no-proof: yes;">17.<span style="mso-tab-count: 1;"> </span>Pittenger MF, Mackay AM, Beck SC, Jaiswal RK, Douglas R, Mosca
JD, Moorman MA, Simonetti DW, Craig S, & Marshak DR (1999) Multilineage
potential of adult human mesenchymal stem cells. <i style="mso-bidi-font-style: normal;">Science</i> 284(5411):143-147. </span></a><span style="mso-no-proof: yes;"><a href="http://www.ncbi.nlm.nih.gov/pubmed/10102814">http://www.ncbi.nlm.nih.gov/pubmed/10102814</a><o:p></o:p></span></span></div>
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Anonymousnoreply@blogger.com0tag:blogger.com,1999:blog-6181073719515632715.post-34603663926508002692016-04-04T10:07:00.000-04:002016-04-04T10:07:11.522-04:00ORS 2016 Annual Meeting Round-Up<div class="MsoNormal">
<i><span style="font-family: "times" , "times new roman" , serif;">A guest post by RoosterBio Travel Award winner, <a href="http://bonassar.research.engineering.cornell.edu/lab-members/katherine-hudson/" target="_blank">Katherine Hudson</a>. </span></i></div>
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<table cellpadding="0" cellspacing="0" class="tr-caption-container" style="float: right; margin-left: 1em; text-align: right;"><tbody>
<tr><td style="text-align: center;"><a href="https://2.bp.blogspot.com/-YQQdzKOd1f8/VwJ0nadME4I/AAAAAAAAD0A/7fH06z0GY_I61ObllNBG5SRIz67_u3a7g/s1600/Katie%2BORS.jpg" imageanchor="1" style="clear: right; margin-bottom: 1em; margin-left: auto; margin-right: auto;"><img border="0" height="320" src="https://2.bp.blogspot.com/-YQQdzKOd1f8/VwJ0nadME4I/AAAAAAAAD0A/7fH06z0GY_I61ObllNBG5SRIz67_u3a7g/s320/Katie%2BORS.jpg" width="320" /></a></td></tr>
<tr><td class="tr-caption" style="text-align: center;"><b>Rocking some RoosterBio swag!</b></td></tr>
</tbody></table>
</div>
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<span style="font-family: "times" , "times new roman" , serif;">The <a href="http://www.ors.org/">Orthopaedic Research
Society</a> (ORS) Annual Meeting brings together clinicians, scientists, and
engineers dedicated to addressing the current challenges facing orthopaedic
research. With <a href="http://www.ors.org/wp-content/uploads/2016/02/ORS-2016-PROGRAM-BOOK_FOR_WEB.pdf">over
2,200 abstracts</a> being presented, it can be a difficult landscape to
navigate. Luckily, the organizers make it easy to connect with researchers with
similar interests while still facilitating expanded horizons. <o:p></o:p></span></div>
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<span style="font-family: "times" , "times new roman" , serif;"><br /></span>
<span style="font-family: "times" , "times new roman" , serif;">My research focuses on tissue engineering of the <a href="https://en.wikipedia.org/wiki/Intervertebral_disc">intervertebral disc</a>
(IVD), using mechanical and chemical cues to encourage <a href="http://www.roosterbio.com/pages/what-are-mscs">Mesenchymal Stem Cell</a>
(MSC) differentiation and tissue maturation within my constructs. This subject
spans the topics of stem cell biology, biomaterial development, and<i> in vivo </i>preclinical trials, making the
ORS a perfect place to present my work. Attending the <a href="http://www.ors.org/2016annualmeeting/">ORS meeting</a> allowed me to
accomplish many things including sharing my most recent work, networking with
potential employers and collaborators, and learning about the latest scientific
developments and techniques. <o:p></o:p></span></div>
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<div class="separator" style="clear: both; text-align: center;">
<a href="https://2.bp.blogspot.com/-Wzgybbznm0w/VwJrW-jpKEI/AAAAAAAADzs/p-yai_6v-2QeDRalgW5prUbN1kl9tj45A/s1600/Hudson%2BIVD.tif" imageanchor="1" style="clear: left; float: left; margin-bottom: 1em; margin-right: 1em;"><img border="0" height="138" src="https://2.bp.blogspot.com/-Wzgybbznm0w/VwJrW-jpKEI/AAAAAAAADzs/p-yai_6v-2QeDRalgW5prUbN1kl9tj45A/s400/Hudson%2BIVD.tif" width="400" /></a></div>
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<span style="font-family: "times" , "times new roman" , serif;">During the conference, my posters received plenty of traffic,
which extended the impact of my findings. Both posters challenge traditional
tissue engineering paradigms, and my aim was to make other tissue engineers
aware of the potential benefits of culturing (and expanding) MSCs in hypoxia,
and immunophenotyping cells before and after their use in 3D scaffolds (See my
ORS abstracts <a href="https://cdn.shopify.com/s/files/1/0515/3253/files/MHCExpression_Hudson.pdf?9731365873884763917">here</a>
and <a href="https://cdn.shopify.com/s/files/1/0515/3253/files/HypoxiaDiscs_RB_Hudson.pdf?18030064184256516333">here</a>
for details). Additionally, I was able to get valuable feedback on my research
that will make my upcoming dissertation stronger. <o:p></o:p></span></div>
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<span style="font-family: "times" , "times new roman" , serif;">The ORS encourages and facilitates networking with both
clinicians and other scientists. While at the conference, I met with
researchers from across the country, and even interviewed for postdoctoral
positions, the next step after I finish my PhD work this May. Through these
discussions and the presentation sessions organized by the ORS, I was exposed
to the latest research in my current and proposed fields of study. This
included the newest cell culture techniques, evaluation tools, and IVD biology.
<o:p></o:p></span></div>
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<span style="font-family: "times" , "times new roman" , serif;"><br /></span></div>
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<span style="font-family: "times" , "times new roman" , serif;">Although I am biased towards tissue engineering and
development, I feel that these topics were the highlight of the ORS meeting
this year. The source of cells used in regenerative therapies, be they primary
or stem cells, was a focus throughout the conference. Additionally, novel
biomaterials and stimulation techniques to drive the behavior of cells was a
focus. It is important that researchers understand the structure of orthopaedic
tissues and their failure modes over multiple scales before we can truly
develop successful repair and regeneration strategies. Appropriate cells types
and materials facilitate these studies.</span></div>
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<span style="font-family: "times" , "times new roman" , serif;">Some presentations that stood out to me:</span></div>
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<span style="font-family: "times" , "times new roman" , serif;"></span><br />
<a name='more'></a></div>
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<b><span style="line-height: 115%;"><span style="font-family: "times" , "times new roman" , serif;">Hyperspectral Raman Imaging as a
Novel Tool for Quantifying the Distribution of Biochemical Constituents in
Native and Engineered Cartilage</span></span></b></div>
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</div>
<ul>
<li><span style="font-family: "times" , "times new roman" , serif;"><span style="line-height: 115%; text-indent: -0.25in;"><span style="font-stretch: normal; line-height: normal;"> </span></span><span style="line-height: 115%; text-indent: -0.25in;">Dr.
Michael Albro, </span><a href="http://www.stevensgroup.org/index.php/site" style="text-indent: -0.25in;"><span style="line-height: 115%;">Stevens Lab</span></a><span style="line-height: 115%; text-indent: -0.25in;">, Imperial College London</span></span></li>
<li><span style="line-height: 115%; text-indent: -0.25in;"><span style="font-family: "times" , "times new roman" , serif;">Hyperspectral
Raman imaging can be used to determine the distribution of extracellular matrix
components (including collagen and glycosaminoglycans) by separating the raw
spectra into its constituents using multivariate curve resolution analysis.
This technique may be used to determine structure function relationships in
native and engineered tissues.</span></span></li>
<li><span style="font-family: "times" , "times new roman" , serif;"><span style="line-height: 115%; text-indent: -0.25in;">A
related publication can be found </span><a href="http://www.ncbi.nlm.nih.gov/pubmed/?term=Heterogeneous+engineered+cartilage+growth+results+from+gradients+of+media-supplemented+active+TGF-%CE%B2+and+is+ameliorated+by+the+alternative+supplementation+of+latent+TGF-%CE%B2" style="text-indent: -0.25in;"><span style="line-height: 115%;">here</span></a><span style="line-height: 115%; text-indent: -0.25in;">.</span></span></li>
</ul>
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<b><span style="line-height: 115%;"><span style="font-family: "times" , "times new roman" , serif;">Expansion of Mesenchymal Stem Cells
on Nanofibrous Scaffolds Preserves Their ‘Naïve’ Mechanobiologic State</span></span></b></div>
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</div>
<ul>
<li><span style="font-family: "times" , "times new roman" , serif;"><span style="line-height: 115%; text-indent: -0.25in;"><span style="font-stretch: normal; line-height: normal;"> </span></span><span style="line-height: 115%; text-indent: -0.25in;">Dr.
Su-Jin Heo, </span><a href="http://www.med.upenn.edu/orl/maucklab/index.shtml" style="text-indent: -0.25in;"><span style="line-height: 115%;">Mauck Lab</span></a><span style="line-height: 115%; text-indent: -0.25in;">, University of Pennsylvania</span></span></li>
<li><span style="font-family: "times" , "times new roman" , serif;"><span style="line-height: 115%; text-indent: -0.25in;"><span style="font-stretch: normal; line-height: normal;"> </span></span><span style="line-height: 115%; text-indent: -0.25in;">Bovine
MSCs cultured on tissue culture plastic, which as a super-physiologic
stiffness, respond to their mechanical environment by moving towards a fibrotic
phenotype. This behavior persists even after the cells are moved to a different
culture substrate, indicating that the cells have ‘mechanical memory’. These
effects were not seen when MSCs were cultured on PCL nanofibrous scaffolds,
indicating that these cells may be retaining their naïve phenotype, including
multipotency.</span></span></li>
<li><span style="font-family: "times" , "times new roman" , serif;"><span style="line-height: 115%; text-indent: -0.25in;">A
related publication can be found </span><a href="http://www.ncbi.nlm.nih.gov/pubmed/?term=Microstructural+heterogeneity+directs+micromechanics+and+mechanobiology+in+native+and+engineered+fibrocartilage" style="text-indent: -0.25in;"><span style="line-height: 115%;">here</span></a><span style="line-height: 115%; text-indent: -0.25in;">.</span></span></li>
</ul>
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<b><span style="line-height: 115%;"><span style="font-family: "times" , "times new roman" , serif;">Hypoxia promotes stable chondrocyte
differentiation of articular cartilage progenitor cells</span></span></b></div>
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</div>
<ul>
<li><span style="font-family: "times" , "times new roman" , serif;"><span style="line-height: 115%; text-indent: -0.25in;"><span style="font-stretch: normal; line-height: normal;"> </span></span><span style="line-height: 115%; text-indent: -0.25in;">Devon
Anderson, </span><a href="http://www.ohsu.edu/xd/health/services/ortho/research-training/research-programs/johnstone-lab.cfm?WT_rank=1" style="text-indent: -0.25in;"><span style="line-height: 115%;">Johnstone Lab</span></a><span style="line-height: 115%; text-indent: -0.25in;">, Oregon Health and Science
University</span></span></li>
<li><span style="font-family: "times" , "times new roman" , serif;"><span style="line-height: 115%; text-indent: -0.25in;"><span style="font-stretch: normal; line-height: normal;"> </span></span><span style="line-height: 115%; text-indent: -0.25in;">Low
oxygen tension (2% oxygen) significantly increased the expression of genes
associated with chondrogenesis including COL2A1, SOX9, and ACAN, while down
regulating expression of COL10A1 and MMP13 compared to ‘normal’ oxygen tension
(20% oxygen) in pellet culture. Significant variation in response was seen
between donors.</span></span></li>
<li><span style="font-family: "times" , "times new roman" , serif;"><span style="line-height: 115%; text-indent: -0.25in;">A
related publication can be found </span><a href="http://www.ncbi.nlm.nih.gov/pmc/articles/PMC3979022/" style="text-indent: -0.25in;"><span style="line-height: 115%;">here</span></a><span style="line-height: 115%; text-indent: -0.25in;">.</span></span></li>
</ul>
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<b><span style="line-height: 115%;"><span style="font-family: "times" , "times new roman" , serif;">A Secretomic Comparison of the
Induction of Chondrogenesis in Human Mesenchymal Stem Cells via TGF-β1 and
Mechanical Load</span></span></b></div>
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</div>
<ul>
<li><span style="font-family: "times" , "times new roman" , serif;"><span style="line-height: 115%; text-indent: -0.25in;">Oliver
Gardner, </span><a href="https://www.aofoundation.org/Structure/research/exploratory-applied-research/research-institute/research-institute-services/musculoskeletal-regeneration/Pages/Martin-Stoddart.aspx" style="text-indent: -0.25in;"><span style="line-height: 115%;">Stoddart Lab</span></a><span style="line-height: 115%; text-indent: -0.25in;">, AO Research Institute Davos</span></span></li>
<li><span style="font-family: "times" , "times new roman" , serif;"><span style="line-height: 115%; text-indent: -0.25in;"><span style="font-stretch: normal; line-height: normal;"> </span></span><span style="line-height: 115%; text-indent: -0.25in;">Human
MSCs that have been stimulated to undergo chondrogenesis through mechanical
loading or exposure to TGF-β1 secrete different proteins associated chondrogenesis.
Specifically, leptin, leptin receptor, and MDC were higher in TGF-β1. Molecules
produced after mechanical loading point to nitric oxide (NO) as a mediator of
chondrogenesis in this case.</span></span></li>
<li><span style="font-family: "times" , "times new roman" , serif;"><span style="line-height: 115%; text-indent: -0.25in;">A
related publication can be found </span><a href="http://www.ncbi.nlm.nih.gov/pubmed/16804949" style="text-indent: -0.25in;"><span style="line-height: 115%;">here</span></a><span style="line-height: 115%; text-indent: -0.25in;">.</span></span></li>
</ul>
<div style="text-indent: -24px;">
<span style="font-family: "times" , "times new roman" , serif;"><span style="line-height: 18.4px;"><br /></span></span></div>
<div style="text-align: left; text-indent: -24px;">
<span style="text-indent: 0px;"><span style="font-family: "times" , "times new roman" , serif;">Overall, the ORS Annual Meeting was a great venue to present my work and learn from the many other dedicated researchers in my and related fields. I am grateful to RoosterBio Inc. not only for the cells I use to complete my research, but also the travel award in support of my attending this conference. </span></span></div>
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Anonymousnoreply@blogger.com5tag:blogger.com,1999:blog-6181073719515632715.post-23777941270590401922016-03-28T12:54:00.001-04:002016-03-28T12:56:48.031-04:00Towards a Cell Therapy Manufacturing Technology Roadmap -- Resources<div class="separator" style="clear: both; text-align: center;">
<a href="http://3.bp.blogspot.com/-qKkBdiGcya0/VvlgR720S8I/AAAAAAAADyk/2rLOcupOuxEz9DFt0bWzhswi7l_0a2kOA/s1600/canstock8171422.jpg" imageanchor="1" style="clear: right; float: right; margin-bottom: 1em; margin-left: 1em;"><img border="0" src="https://3.bp.blogspot.com/-qKkBdiGcya0/VvlgR720S8I/AAAAAAAADyk/2rLOcupOuxEz9DFt0bWzhswi7l_0a2kOA/s400/canstock8171422.jpg" /></a></div>
<span style="font-family: "arial" , "helvetica" , sans-serif;">RoosterBio was founded to accelerate the development of the Regenerative Medicine field by utilizing advancements in Manufacturing Sciences. As such, we have long been involved in laying the groundwork for a Technology Roadmap for sustainable Cell Therapy Manufacturing. </span><br />
<span style="font-family: "arial" , "helvetica" , sans-serif;"><br /></span>
<span style="font-family: "arial" , "helvetica" , sans-serif;">Recently, RoosterBio was part of the <a href="http://www.nist.gov/amo/amtech/70nanb14h048.cfm" target="_blank">first-ever Cell Manufacturing Consortium</a> aiming to position the United States as a leading developer of Cell Manufacturing Technologies and the Chief Authority on Cell Manufacturing Standards Worldwide. The goal of this Consortium was to e<span style="background-color: white; line-height: 19.2px;">stablish a collaborative public-private partnership that engages industry, academia, regulators, and other stakeholders in removing barriers to the advancement of the cell-manufacturing industry, thereby bringing new therapies and diagnostics to the healthcare market. </span> As a result, the Consortium members have formulated an extensive Cell Therapy Manufacturing Technology Roadmap to the year 2025. This document is in final review and should be available shortly. In the meantime, we've compiled some resources (to which we will continue to add and seek your input as well) on Cell Therapy Manufacturing for those interested.</span><br />
<span style="font-family: "arial" , "helvetica" , sans-serif;"><u><br /></u></span>
<b><span style="font-family: "arial" , "helvetica" , sans-serif;"><u>Relevant Publications:</u></span></b><br />
<span style="color: blue;"><br /></span>
<ul>
<li><a href="http://www.aiche.org/resources/publications/cep/2010/november/developing-cell-therapy-biomanufacturing-processes" target="_blank"><span style="color: blue; font-family: "arial" , "helvetica" , sans-serif;">Developing Cell Therapy Biomanufacturing Processes</span></a></li>
<li><a href="http://www.bioprocessintl.com/manufacturing/cell-therapies/cell-therapy-bioprocessing-314870/" target="_blank"><span style="color: blue; font-family: "arial" , "helvetica" , sans-serif;">Cell Therapy Bioprocessing</span></a></li>
<li><span style="color: blue; font-family: "arial" , "helvetica" , sans-serif;"><a href="http://www.bioprocessintl.com/manufacturing/cell-therapies/meeting-lot-size-challenges-of-manufacturing-adherent-cells-for-therapy-328093/" target="_blank">Meeting Lot-Size Challenges of Manufacturing Adherent Cells for Therapy</a></span></li>
<li><a href="http://www.ncbi.nlm.nih.gov/pubmed/22168500" style="background-color: white; font-family: Arial, Helvetica, sans-serif; line-height: 1.125em;" target="_blank"><span style="color: blue;">Developing assays to address identity, potency, purity and safety: cell characterization in cell therapy process development.</span></a></li>
</ul>
Anonymousnoreply@blogger.com2tag:blogger.com,1999:blog-6181073719515632715.post-15402021388205003052016-02-23T12:36:00.002-05:002016-02-24T16:08:18.830-05:00At the Cutting-Edge of Regenerative Medicine: Bioengineering Human-Sized Bone<style>
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<tr><td style="text-align: center;"><a href="https://1.bp.blogspot.com/-kwuiGjfT1_E/VsyX-xGZigI/AAAAAAAADwk/PAWGvHXC0A0/s1600/A00521F01.jpg" imageanchor="1" style="clear: right; margin-bottom: 1em; margin-left: auto; margin-right: auto;"><img border="0" height="320" src="https://1.bp.blogspot.com/-kwuiGjfT1_E/VsyX-xGZigI/AAAAAAAADwk/PAWGvHXC0A0/s320/A00521F01.jpg" width="203" /></a></td></tr>
<tr><td class="tr-caption" style="text-align: center;"><div style="text-align: left;">
Fisher et al published the first attempt at engineering</div>
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a full-scale adult human femur head from hMSCs. This</div>
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is the largest reported tissue to have been engineered</div>
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and took over 700 million hMSCs to fabricate.</div>
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<a href="http://teblumd.net/"><span style="font-family: "arial";">John
Fisher’s laboratory</span></a><span style="font-family: "arial";"> at the <span style="mso-no-proof: yes;">University of Maryland, College Park recently
published what can be considered</span> a significant advance for Tissue
Engineering and Regenerative Medicine. <span style="mso-no-proof: yes;">Graduate
student Bao Nguyen and her colleagues have engineered a </span></span><a href="http://www.ncbi.nlm.nih.gov/pubmed/26653703"><span style="font-family: "arial"; mso-no-proof: yes;">bone construct that is 20 times larger than any
reported previously</span></a><span style="font-family: "arial"; mso-no-proof: yes;">,
and the size of an adult human femur.</span></div>
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<span style="font-family: "arial"; mso-no-proof: yes;">Why is this important?
Critical size bone defects are a significant health problem (resulting in over
$1 billion in annual healthcare costs incurred in the U.S.) and are currently
treated with grafts, decellularized bone, or synthetic bone grafts, with
sometimes unsucessful results. As such, modern medicine has been looking to tissue
engineered bone grafts as future treatments for such defects. </span><a href="http://roosterbio.blogspot.com/2014/03/mesenchymal-stem-cells-workhorse-of.html"><span style="font-family: "arial"; mso-no-proof: yes;">Human bone marrow-derived Mesenchymal
Stem Cells</span></a><span style="font-family: "arial"; mso-no-proof: yes;"> (hBM-MSCs)
are a promising cell source for such applications because they </span><a href="http://roosterbio.blogspot.com/2014/05/mscs-as-workhorse-of-regenerative.html"><span style="font-family: "arial"; mso-no-proof: yes;">efficiently differentiate down the
osteogenic path</span></a><span style="font-family: "arial"; mso-no-proof: yes;"> and
also </span><a href="http://roosterbio.blogspot.com/2014/12/priming-of-hmscs-to-improve-potency.html"><span style="font-family: "arial"; mso-no-proof: yes;">secrete paracrine factors</span></a><span style="font-family: "arial"; mso-no-proof: yes;"> that may aid survival and
vascularization of engineered bone. Prior to this publication, engineered
constructs have been relatively small due to cell and culture limits.<span style="mso-spacerun: yes;"> </span>One major challenge has been growing
hBM-MSCs, while maintaining their function, to sufficient numbers needed for an
adult human-sized construct; a </span><a href="http://www.roosterbio.com/products/msc001"><span style="font-family: "arial"; mso-no-proof: yes;">challenge </span></a><span style="font-family: "arial"; mso-no-proof: yes;">adressed by RoosterBio.<span style="mso-spacerun: yes;">
</span>In addition, nutrient and O<sub>2</sub> transfer are often insufficient
to maintain cell viability and function throughout larger constructs,
especially those of adult human dimension.</span></div>
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<span style="font-family: "arial"; mso-no-proof: yes;">To address this cell
culture limitation, the Fisher laboratory developed a T</span><span style="font-family: "arial";">ubular Perfusion System (TPS) bioreactor where cells
and scaffolds are cultured in a cylindrical chamber and subject to circular media
flow. This system has high nutrient and O<sub>2</sub> transfer and efficient waste
removal and has been previously used to produce smaller engineered bone and
cartilage constructs (See </span><a href="http://www.ncbi.nlm.nih.gov/pubmed/26724678"><span style="font-family: "arial";">here</span></a><span style="font-family: "arial";"> and </span><a href="http://www.ncbi.nlm.nih.gov/pubmed/26577256"><span style="font-family: "arial";">here</span></a><span style="font-family: "arial";">).</span></div>
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<span style="font-family: "arial"; mso-no-proof: yes;">In the study detailed
here, the authors had access to and combined, for the first time, advanced
technologies required for </span><a href="http://www.roosterbio.blogspot.com/2015/01/the-rise-of-biofabrication-and.html"><span style="font-family: "arial"; mso-no-proof: yes;">biofabrication</span></a><span style="font-family: "arial"; mso-no-proof: yes;">: 3D printing, the TPS bioreactor,
and scalable production of hBM-MSCs. The goal of the study was to scale-up bone
constructs to adult human size. A full size mold of the superior portion of a
human femur (the largest bone in the human body) was 3D printed using
information from an </span><a href="https://grabcad.com/"><span style="font-family: "arial"; mso-no-proof: yes;">opensource database</span></a><span style="font-family: "arial"; mso-no-proof: yes;">. The mold measured 23 cm long and
10 cm at its widest point with a volume of<span style="mso-spacerun: yes;">
</span>200 cm<sup>3</sup>. The mold was filled with hBM-MSCs in alginate beads
(3 mm beads, 100,000 cells per bead). The entire construct utlillized 7200
aliginate beads containing a total of 7.2 x 10<sup>8</sup> cells (yes, that is
720 million cells!). The high volume hBM-MSC cell and media systems used were
from RoosterBio, and technical support for the efficient production
of large volumes of hBM-MSCs was provided by our company.</span></div>
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<br /></div>
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<span style="font-family: "arial"; mso-no-proof: yes;">After 8 days of
culture in the TPS, the construct was examined for cell viability and bone
differentiation. High cell viability was seen in all parts of the construct,
both on the outside and the inside (interior). In addition, hBM-MSCs committed
to the osteogenic lineage throughout the construct, demonstrating efficiency of the TPS
culture system. Both early (Alkaline Phosphotase, ALP) and late (Bone
Morphogenic Protein-2, BMP-2) markers of osteogenesis were upregulated relative
to day 0. Interestingly, ALP and BMP-2 expression was 25- to 30-fold higher in
the construct shaft relative to other portions of the construct. The authors
speculate that this is due to shear stress exerted on the parts of the
construct closest to the inlet, which activates hBM-MSC signaling pathways,
causing release of paracrine factors that stimulate osteogenesis of the
“downstream” shaft portion. Taken together, these results demonstrate that the
confluence of cutting-edge technologies such as 3D printing, TPS bioreactors,
and best-in-class hBM-MSC manufacturing processes enable the engineering of
adult human-sized tissue constructs.</span></div>
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<br /></div>
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<span style="font-family: "arial"; mso-no-proof: yes;">While “…this first
foray into full-scale bone engineering provides the foundation for future
clinical applications of bioengineered bone grafts…” the authors point out some
limitations to this study. The 8 day culture period was relatively short, given
the weeks usually needed for high efficiency bone differentiation. Thus,
extended time points and the fabrication of additional large constructs are
needed to fully explore the capabilities of the TPS system. Further, alginate is
a soft material, and its mechanical properties do not render it the best suited
for bone differentiation.<span style="mso-spacerun: yes;"> </span>In addition,
hBM-MSCs within aliginate beads lack cell:cell contact, which may also limit their
osteogenic differentiation.<span style="mso-spacerun: yes;"> </span>To address
these limitations of the current system, the Fisher group is developing a 3D
printed shell made of an implantable rigid material better suited for the
engineering of bone constructs. <span style="mso-spacerun: yes;"> </span>Finally,
the construct lacks a vascular network, which can be overcome by including
endothelial cells (EC) in addition to hBM-MSCs or by incorporating
micro-channels in the engineered constructs through a variety of methods (e.g. biomaterial
fabrication and 3D printing). Despite the aforementioned limitations, the
work presented is a significant advance towards clinical-sized tissue-engineered
bone constructs for use in patients.</span></div>
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<br /></div>
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<div style="text-align: justify;">
<span style="font-family: "arial"; mso-no-proof: yes;">In an attempt to
elicit discussion, I will mention other methods that harbor potential for use
in such large-scale tissue engineering applications. For one, hBM-MSC
aggregates could be used in place of cells in alginate beads. These </span><a href="http://roosterbio.blogspot.com/2014/10/rapid-and-economic-generation-of-hmsc.html"><span style="font-family: "arial"; mso-no-proof: yes;">3D-MSC</span></a><span style="font-family: "arial"; mso-no-proof: yes;"> not only maintain cell:cell contact
but also </span><a href="http://www.ncbi.nlm.nih.gov/pubmed/21833761"><span style="font-family: "arial"; mso-no-proof: yes;">undergo osteogenic differentiation
more efficiently</span></a><span style="font-family: "arial"; mso-no-proof: yes;">
than cells grown on tissue culture plastic, are resistant to </span><a href="http://www.ncbi.nlm.nih.gov/pubmed/24168395"><span style="font-family: "arial"; mso-no-proof: yes;">hypoxia</span></a><span style="font-family: "arial"; mso-no-proof: yes;">, and secrete angiogenic cytokines. Secondly, factors that
stimulate bone differentiation of hBM-MSCs, and/or alter mechanical properties
of the construct, could be incorporated into the polymer scaffolding, or could
be </span><a href="http://www.ncbi.nlm.nih.gov/pubmed/22658155"><span style="font-family: "arial"; mso-no-proof: yes;">introduced into 3D-MSC aggregates</span></a><span style="font-family: "arial"; mso-no-proof: yes;">. Finally, once bio-inks are
developed further, the bone construct could be patterned by 3D printing of cells
(hBM-MSC, EC, 3D-MSC) and materials.<span style="mso-spacerun: yes;"> </span>Now
that human-sized constructs are possible in terms of cell numbers and O<sub>2</sub>
and nutrient diffusion, the possibilities are virtually endless.</span></div>
</div>
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<div style="text-align: justify;">
<br /></div>
</div>
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<div style="text-align: justify;">
<span style="font-family: "arial"; mso-no-proof: yes;">Finally, we sincerely
thank Bao Nguyen and John Fisher for being early adopters of RoosterBio hBM-MSCs
and joining us in accelerating Regenerative Medicine!</span></div>
</div>
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<div style="text-align: justify;">
<br /></div>
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<b style="mso-bidi-font-weight: normal;"><span style="font-family: "arial";">References:</span></b></div>
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<br /></div>
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<span style="font-family: "arial";">Nguyen BB, Ko H, Moriarty RA, Etheridge
JM, Fisher JP. Dynamic Bioreactor Culture of High Volume Engineered Bone
Tissue. Tissue Engineering Part A. Volume 22, Numbers 3 and 4, 2016, ahead of
print. doi:10.1089/ten.tea.2015.0395. <span style="mso-spacerun: yes;"> </span></span><a href="http://online.liebertpub.com/doi/abs/10.1089/ten.tea.2015.0395"><span style="font-family: "arial";">http://online.liebertpub.com/doi/abs/10.1089/ten.tea.2015.0395</span></a><span style="font-family: "arial";"><span style="mso-spacerun: yes;"> </span></span></div>
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<span style="font-family: "arial";">I’m sorry that this is paywalled!</span></div>
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<br /></div>
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<span style="font-family: "arial";">Yeatts, A.B., and Fisher, J.P. Tubular
perfusion system for the long-term dynamic culture of human mesenchymal stem
cells. Tissue Eng Part C 17, 337, 2011. </span></div>
<div class="MsoNormal" style="line-height: normal; margin-bottom: .0001pt; margin-bottom: 0in;">
<a href="http://online.liebertpub.com/doi/abs/10.1089/ten.tec.2011.0263"><span style="font-family: "arial";">http://online.liebertpub.com/doi/abs/10.1089/ten.tec.2011.0263</span></a><span style="font-family: "arial";"></span></div>
<div class="MsoNormal" style="line-height: normal; margin-bottom: .0001pt; margin-bottom: 0in;">
<br /></div>
<div class="MsoNormal" style="line-height: normal; margin-bottom: .0001pt; margin-bottom: 0in;">
<span style="font-family: "arial";">Yeatts, A.B., Choquette, D.T., and
Fisher, J.P. Bioreactors to influence stem cell fate: augmentation of
mesenchymal stem cell signaling pathways via dynamic culture systems. Biochim
Biophys Acta 1830, 2470, 2013. </span></div>
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<a href="http://www.ncbi.nlm.nih.gov/pmc/articles/PMC3461086/"><span style="font-family: "arial";">http://www.ncbi.nlm.nih.gov/pmc/articles/PMC3461086/</span></a><span style="font-family: "arial";"> </span></div>
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<br /></div>
<div class="MsoNormal" style="line-height: normal; margin-bottom: .0001pt; margin-bottom: 0in;">
<span style="font-family: "arial"; mso-no-proof: yes;">Ma, X et al. Deterministically
patterned biomimetic human iPSC-derived hepatic model via rapid 3D bioprinting<span style="mso-spacerun: yes;"> </span>PNAS, Early Edition doi:
10.1073/pnas.1524510113 </span><a href="http://www.pnas.org/content/early/2016/02/04/1524510113"><span style="font-family: "arial"; mso-no-proof: yes;">http://www.pnas.org/content/early/2016/02/04/1524510113</span></a><span style="font-family: "arial"; mso-no-proof: yes;"> </span></div>
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Anonymousnoreply@blogger.com1tag:blogger.com,1999:blog-6181073719515632715.post-844327469427993652016-02-12T22:59:00.002-05:002016-02-12T22:59:34.930-05:00<div class="MsoNormal" style="background: white;">
<span style="color: #222222; font-family: "arial" , sans-serif; font-size: 12.0pt;">Dear Stem Cell Pioneers:<o:p></o:p></span></div>
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<span style="color: #222222; font-family: "arial" , sans-serif; font-size: 12.0pt;"><br /></span></div>
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<span style="color: #222222; font-family: "arial" , sans-serif; font-size: 12.0pt;"> I am as excited as ever about Cell Therapy and Regenerative
Medicine, and the role that RoosterBio is playing in advancing these Industries. Each February marks the anniversary of when
we shipped our first stem cell products – 2 years now on the market! We have shipped our products to 13
countries worldwide, across 5 different continents, and are continuing to
expand our reach in the stem cell marketplace.
As the CEO of RoosterBio, I like to take time to reflect during these
milestones on where we have been, where we currently are, and where we are
going. This is important to make sure
that we are still being true to the mission of the company, the vision that we
hold for the industry, and to course correct if necessary. This keeps us on our trajectory of helping to build a sustainable, lasting business that is helping you, our
customers, do amazing things with living cells. </span></div>
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<span style="color: #222222; font-family: "arial" , sans-serif; font-size: 12.0pt;"> Looking forward into 2016 and beyond, our mission is still intact:
to greatly increase the availability and accessibility of stem cell technologies
to researchers and product developers across the globe. We remain firmly committed to our vision of
accelerating the pace of product development and clinical translation in
cellular therapy, bioprinting and tissue engineering. We have made great strides in 2015 towards
these goals, and 2016 brings great opportunity as we continue our journey.<o:p></o:p></span></div>
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<span style="color: #222222; font-family: "arial" , sans-serif; font-size: 12.0pt;"> In 2015, we built upon our hMSC technology platform to launch
products focused on two key areas critical to the future of Regenerative
Medicine: bioprinting and stem cell manufacturing sciences. We launched several unique First-In-Class products that greatly simplify R&D in these critical stem cell sub-disciplines,
lowering barriers and accelerating discovery and eventual commercial
translation. These unprecedented products include:<o:p></o:p></span></div>
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<li><span style="color: #222222; font-family: "arial" , sans-serif; font-size: 12pt; text-indent: -0.25in;"><a href="http://www.prweb.com/releases/2015/10/prweb13027227.htm">RTP</a>
(Ready-to-Print) cells, the first stem cell reagent</span></li>
<li><span style="color: #222222; font-family: "arial" , sans-serif; font-size: 12pt; text-indent: -0.25in;"><a href="http://roosterbio.blogspot.com/2015/08/enabling-new-paradigm-in-hmsc.html">RoosterReplenish</a>,
the first hMSC suspension bioreactor media feed</span></li>
<li><span style="color: #222222; font-family: "arial" , sans-serif; font-size: 12pt; text-indent: -0.25in;"><a href="http://www.prweb.com/releases/2015/09/prweb12988213.htm">Bioreactor
kits</a> and accompanying protocols to simplify and streamline scalable
suspension bioreactor expansion of MSCs</span></li>
<li><span style="color: #222222; font-family: "arial" , sans-serif; font-size: 12pt; text-indent: -0.25in;"><a href="http://3dprintingindustry.com/2015/09/25/cellink-launches-first-commercial-living-cellular-consumables-bioprinting/">Bioprinting
kits</a> and accompanying protocols for simplified mixing and printing of
living cellular bioinks</span></li>
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<span style="color: #222222; font-family: "arial" , sans-serif; font-size: 12.0pt;"> Our customers have also been very busy working on their own
innovations, and the last year has brought <a href="http://www.roosterbio.com/pages/the-brainy-stuff">multiple abstracts</a>
presented at international conferences (including the <a href="http://www.ors.org/">ORS</a>, <a href="http://www.celltherapysociety.org/">ISCT</a>
and <a href="http://www.isscr.org/">ISSCR</a> Annual Meetings and the <a href="http://www.termis.org/">TERMIS World Congress</a>), the first
publications using our cells for <a href="http://www.ncbi.nlm.nih.gov/pubmed/26139547">bioreactor expansion</a> and
<a href="http://online.liebertpub.com/doi/abs/10.1089/ten.tea.2015.0395">engineering
massive tissue constructs</a>, as well as numerous grant proposals that have been submitted using our products as key reagents. <o:p></o:p></span></div>
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<span style="color: #222222; font-family: "arial" , sans-serif; font-size: 12.0pt;"> The government’s interest in Therapeutic Cell Manufacturing
Sciences has also expanded, and it is accepted now that the US must invest in
these technologies and build our own infrastructure and talent base in this
critical field (see <a href="http://www.nist.gov/amo/nnmi/upload/NIST-NNMI-NOI-2015-12-21.pdf">here</a>
and <a href="https://www.fbo.gov/index?s=opportunity&mode=form&id=414582ce17ec70815d488af87a42090b&tab=core&_cview=1">here</a>). We had the pleasure of participating on the
executive committee of a NIST-funded <a href="http://www.news.gatech.edu/2016/01/18/center-will-develop-consistent-manufacturing-processes-cell-based-therapies">Cell
Manufacturing Consortium</a> with many academic and industry leaders in the
field of Therapeutic Cell Manufacturing. The output of this is a Technology
Roadmap for Scalable Cell Therapy Manufacturing that will drive funding areas
in this field for years to come (link to be added as soon as the Roadmap is
finalized and published).<o:p></o:p></span></div>
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<span style="color: #222222; font-family: "arial" , sans-serif; font-size: 12.0pt;"> None of this would be possible without the hard work and
dedication of the entire RoosterBio team (and we have already added to the team
in 2016!), as well as the support that we are getting from you, our valued customers. Please continue to join us on our journey as
we accelerate the development of the cell-based <a href="https://www.whitehouse.gov/sites/default/files/microsites/ostp/national_bioeconomy_blueprint_april_2012.pdf">BioEconomy</a>.<o:p></o:p></span></div>
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<span style="color: #222222; font-family: "arial" , sans-serif; font-size: 12.0pt;">All the Best from Frederick, Maryland.</span><span style="color: #222222; font-family: "arial" , sans-serif; font-size: 12pt;"> </span></div>
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<span style="color: #222222; font-family: "arial" , sans-serif; font-size: 12.0pt;">Jon A Rowley<o:p></o:p></span></div>
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<span style="color: #222222; font-family: "arial" , sans-serif; font-size: 12.0pt;">Chief Executive and Technology Officer<o:p></o:p></span></div>
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<span style="color: #222222; font-family: "arial" , sans-serif; font-size: 12.0pt; line-height: 107%;">RoosterBio, Inc</span><span style="color: #222222; font-family: "arial" , sans-serif; font-size: 9.5pt; line-height: 107%;">.</span><o:p></o:p></div>
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