December 15, 2018

Building Effective Multi-Year Process Development Programs: Evolution of Technology Platform Decisions Based on Lot Size

Authored by Josephine Lembong, Ph.D., Scientist, Product & Process Development & Jon Rowley, Ph.D., Founder and Chief Product Officer, RoosterBio Inc.

Previously, we published a blog post on estimating hMSC lot sizes for clinical manufacturing, 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.

“It is the technologist’s and engineer’s jobs to drive the technology platform decision making process”

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 RegenMed 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.

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.

Each phase of a clinical trial is associated with a specific production scale, which dictates the production platform

At the end of the last post, 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 here) for each therapeutic program.

November 17, 2018

RoosterBio Products Continue to Expand Presence in Major Mesenchymal Stem/Stromal Cell (MSC)-related Regenerative Medicine Peer Reviewed Publications

Authored by Joseph Candiello, Ph.D., Technical Application Specialist

Figure 1. Current and emerging applications using hMSCs as a critical raw material.
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 scientific publications and a parallel increased presence in clinical applications over the past 10+ years.  Specific to scientific publications, from 2011 to 2015 there were over 17,000 MSC related publications, 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.

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 “brainy stuff”.  The initial articles using RoosterBio technology were published in Biomacromolecules and Cytotherapy 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 PNAS (twice), Biomaterials, Stem Cells and Translational Medicine, American Journal of Respiratory and Critical Care Medicine, and Nucleic Acids Research.

Figure 2: Key Metrics of Publications using RoosterBio products.
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. 

The breadth of topics included in these publications is as diverse as current applications across the hMSC Regenerative Medicine space – and is called out in our databaseof current RoosterBio product publications. As frequency of publications increases, we will highlight some of these – starting with some recent exciting work being reported across the industry:

September 12, 2018

Building Effective Multi-Year Process Development Programs: Estimating hMSC Lot Size Ranges for Clinical Manufacturing through Commercial Demand – it is all about the assumptions

Authored by Josephine Lembong, Ph.D., Scientist, Product & Process Development & Jon A. Rowley, Ph.D., Founder and Chief Product Officer, RoosterBio Inc.

Jon Rowley, RoosterBio’s Founder and Chief Product Officer, gave a talk at ISCT 2018 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.  

The concept is consistent with all good strategic planning; 
  1. Understand what the future looks like and work backwards,
  2. Create a model using reasonable to conservative assumptions along the way to estimate a range of needs,
  3. 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.
This is the type of consultation that we provide in our Process Design and Acceleration business unit at RoosterBio, which we offer to our customers that are incorporating RoosterBio hMSC cell banks and bioprocess media systems 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.

Notes to readers: this exercise has the base assumption that this is an Allogenic, Universal Donor manufacturing process [1] 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.

Lot Size Targets Dictate the Production Platform 

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.

First Start with the End in Mind – Engage with Marketing

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 [2]. 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.

“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”

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.

Dosing Assumptions: this is one of the hardest parts

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 [2,3]. 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 [4,5]. In this blog post we will perform all calculations with an assumption of 50 million cells per dose.

Calculating Yearly Product Needs at Peak Commercial Demand
  • At peak demand, we aim to produce 100,000 doses per year, with a size of 50 million cells per dose.
  • 100,000 doses per year ≈ 2,000 doses per week
    • 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)
    • 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
    • 100,000 doses per year / ~23 lots per year = 4,450 doses/lot 
    • Assume 10% of lot goes to testing (~450 dose), so a final target lot size of 4,900 doses/lot will be needed to be manufactured at peak commercial demand
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 Regenerative Medicine. We will create estimates for peak demand, but then focus on building a reasonable “go-to-market” lot size and manufacturing process.

Calculating Total Cells to Manufacture that are Required to Fill into the Final Container
  • We will assume 1 dose goes into one container.
  • We assumed a mid-range dose size of 50 million (i.e. 50 million viable cells per vial, post-thaw).
  • Cell Count 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.
    • 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).
    • Based on the above assumptions, to consistently achieve 50 million viable cells after thaw, it is required to target filling at 75 million viable cells into the final container:
      • 75 million viable cells – 15 million (20% total cell loss on recovery) = 60 million viable cells 
      • 60 million – 9 million (15% viability drop) = 51 million viable cell target specification
  • 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.

Calculating Lot Size Requirements at Peak Commercial Demand

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:
  • 75 million cells/dose × 4900 doses per manufacturing lot = 368 billion viable cells needed to target 4900 doses from every manufacturing lot.
  • We assume 30% post-harvest cell loss during processing for this exercise – thus at least 525 billion cells need to be manufactured in order to have 368 billion cells during the fill unit operation.
    • 525 billion viable cells – 157 billion (30% post-harvest cell loss) = 368 billion viable cells 
  • 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.
In conclusion, we arrived at the following numbers:
  • 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. 
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.  

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.

We’d like to summarize this post with a few number recommendations:

Phase III/Go-to-Market: ~100B cells
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.
Phase I (small scale): 5-10B cells

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.

  1. 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
  2. Olsen et al., 2018. Peak MSC—Are We There Yet? Front Med 5:178. doi: 10.3389/fmed.2018.00178
  3. Squillaro et al., 2016. Clinical Trials With Mesenchymal Stem Cells: An Update. Cell Transplant 25(5):829-848. doi: 10.3727/096368915X689622
  4. 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
  5. 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

May 16, 2018

ISCT 2018: MSC Biomanufacturing, Bioprocessing, Scale-Up, Analytics and Exosome Production Take Center Stage

Authored by Katrina Adlerz, Ph.D. Scientist, Analytical Development, RoosterBio Inc.

RoosterBio attended the International Society for Cell Therapy 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.

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. Jon Rowley, founder and CTO of RoosterBio, also gave two talks discussing innovations to accelerate MSC Biomanufacturing, Bioprocessing and Scale-Up. He shared insights from RoosterBio’s path to GMP-manufactured cells and media as well as strategies for overcoming obstacles in his talk “Technologies for Radically Reducing Development Timelines of hMSC-based Therapeutic Products” during the Strategies for Commercialization Session. (New to MSCs? Read more about MSCs and biomanufacturing here, herehere and here.) For a copy of Jon's talk, email us.

RoosterBio Analytical, Process & Product Development efforts were well-represented with three posters.

- “A Xeno-Free Fed-Batch Microcarrier Suspension Bioreactor System for the Scalable and Economic Expansion of hBM-MSCs” showed that MSC critical quality attributes were maintained in 0.1L and 3L bioreactor culture. Poster.
- “Scalable Xeno-Free Manufacturing of Extracellular Vesicles Derived from Human Mesenchymal/Stromal Stem Cells” explained a process for generating a high yield of EVs/Exosomes from MSCs in a shortened time frame using RoosterNourishTM-MSC-XF Media. Poster.
- “Development & Technology Transfer of a cGMP Potency Assay: Testing of an Ancillary Material for Stem Cell Manufacturing” outlined the steps in assay development, assay qualification/validation, and tech transfer for a custom potency assay based on cell expansion. Poster.
- In addition, the RoosterBio , BioLife Solutions and Brooks Life Science teams collaboratively presented a poster on MSC cryopreservation: "The Effect of Cryomedia Selection and Transient Warming Events on Post-Cryopreservation Human MSC Function". Poster. 

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.

The conference kicked off with a RoosterBio-sponsored workshop: Improving Mesenchymal Stem Cell Potency and Survival. Steven Bauer, Branch Chief of the Cellular and Tissue Therapy Branch  of the US FDA, and Head of the FDA's MSC Consortium, 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 here.  There were also a number of talks discussing clinical trials results. A common theme of the session was the need to develop analytical methods and assays that can predict MSC-based treatment efficacy in patients.

June 29, 2017

Good Manufacturing Practice for Cell and Cell-Based Therapies: Facilities & Quality Control

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 current Good Manufacturing Practice requirements.  (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 Center for Biologics Evaluation and Research (CBER), a division of the U.S. Food and Drug Administration (FDA).

When manufacturing cell and cell-based products, a production facility 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, closed systems and single-use disposables should be used whenever possible to minimize risk of contamination.  Therefore, a cGMP manufacturing facility must include clean rooms 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 21CFR§211 and 21CFR§1271.

As mentioned above, stringent Quality Control systems must be in place to qualify all reagents and processes and to institute Standard Operating Procedures (SOPs) to ensure quality and consistency in manufacturing processes and the end product. 

Facility and Quality Control considerations for cGMP cell manufacturing.

June 23, 2017

Demystifying Clinical Translation of Stem Cell-Based Therapies

As our company matures and we continue to execute to our mission of accelerating Regenerative Medicine, 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 hMSCs and hMSC-based therapeutics.  While the US FDA and other global regulatory agencies provide guidances 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.
How do you move from pre-clinical to clinical studies?
How must you manufacture your cells?
What data do you need to present to the FDA to move to clinical trials?
What sort of characterization assays are needed? 
What is the process for engaging the FDA and moving to clinical trials?
When do I need to do all of this?!

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.

What other questions do you have around clinical translation of hMSCs and hMSC-based therapies?

Check back in next week for our first blog post in this series.

June 15, 2017

Opportunities for BioManufacturing Sciences to Accelerate Upscaled Mesenchymal Stem Cell Manufacturing Technologies

Contributed by: Timothy R. Olsen, PhD - Sr. Scientist, Process and Product Development at RoosterBio Inc.

After the National Institute for Standards and Technology (NIST) identified upcoming challenges in the U.S. biopharmaceutical manufacturing landscape (see document here), the National Institute for Innovation in Manufacturing Biopharmaceuticals (NIIMBL) 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.
Confluent hMSCs on Solohill microcarriers.
Image from RoosterBio Inc
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,” Greg Russotti (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 blockbuster pharmaceuticals. 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 3D microcarrier-based bioreactor systems, but there are still many manufacturing innovations required before these manufacturing platforms can support a commercial cell therapy product.  
The technology gaps that he specifically mentioned for upscaled hMSC manufacturing were downstream processing technologies, specifically the unit operations related to:

April 14, 2017

Orthopaedic Research Spotlight: ORS 2017 Annual Meeting

A guest blog post by RoosterBio Travel Award winner, Poonam Sharma.

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 ORS 2017 an engaging and dynamic conference to attend.

Knockdown of vimentin may affect chondrogenic
extracellular matrix deposition in high density pellets.
shVim-vimentinknockdown, shLacZ-control.
My research in Dr. Adam H. Hsieh’s Orthopaedic Mechanobiology Lab at the University of Maryland centers on the role of vimentin intermediate filaments in governing mesenchymal stem cell (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 here). 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. MSCs from RoosterBio 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.

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.   
One of the most engaging sessions in this conference was a really fun debate about the related futures of regenerative medicine and orthopaedic implants – Will Regenerative Medicine Make Orthopaedic Implants Obsolete in Our Time? It was captivating to hear the discussion about two research and clinical areas that continue to intersect and diverge. Also, the keynote by Dr. Jennifer Doudna summarizing the CRISPR technology that she helped develop was a great overview and the brief discussion about the ethics of gene therapy was thought-provoking.

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:

January 3, 2017

Guest Blog Post: The Ambitious Future of Tissue Engineering

Bagrat Grigoryan and Jordan MillerPhysiologic Systems Engineering & Advanced Materials Lab at Rice University.

Over the last several decades, various advances in tissue engineering have allowed for the not so distant possibility of replacing, repairing, or regenerating injured tissues1. Significant progress has been made in understanding cellular biology as well as pathophysiology and healthy states of tissues. Additionally, a suite of diverse biofabrication technologies and biomaterials has been conceived, enabling fabrication of complex 3D tissues with greater physiological relevance compared to the traditional 2D context that cells are studied in2. 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.

While different fabrication techniques and biomaterials have been explored in fabricating tissues in vitro, 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 potential3. 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 trials4. With over 450 mesenchymal stem cell (MSC)-based trials alone currently ongoing or completed, the regenerative and immunoregulatory properties of MSCs are constantly being exploited to improve the quality of human life5.

Due to the immense therapeutic potential of MSCs, there is a need to rapidly and reproducibly grow a vast amount of MSCs for clinical and research purposes. Although MSCs were identified and isolated from bone marrow more than 40 years ago, we still have not fully mapped their biological characteristics3.

June 16, 2016

At the Cutting Edge of 3D Bioprinting: WBC 2016 Round Up Part II

A guest blog post by RoosterBio Travel Award winner, Ian Kinstlinger.

Presenting on Open-source Selective Laser 
Sintering at WBC2016!
Since 1980, the international community of biomaterials scientists and engineers has convened every four years to discuss the cutting edge of biomaterials research. This year’s 10th 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 Miller Lab at Rice University in both a podium talk and a poster session.

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.

An overview of one method our lab has introduced to create 
embedded vascular networks in biomaterials.
I used my poster to spread the word about our lab’s Open-source Selective Laser Sintering 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 published earlier this year – and RoosterBio hMSCs were absolutely central. Their high quality and robust differentiation response made characterizing biocompatibility of materials quite straightforward.

A couple of key presentations stood out at WBC 2016:

Nano- and Micro-fabricated Hydrogels for Regenerative Engineering
  • Dr. Ali Khademhosseini, Khademhosseini Lab, Harvard University
  •  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.

Injection of Dual-Crosslinking Hydrogels to Limit Infarct Induced Left Ventricular Remodeling
  • Dr. Jason Burdick, Polymeric Biomaterials Laboratory, University of Pennsylvania
  •  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.

Photoreversible patterning of hydrogel biomaterials with site-specifically-modified proteins
  • Dr. Cole DeForest,  DeForest Research Group, University of Washington
  •  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.

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 printers and inks now hitting the market, including RoosterBio’s new ready-to-print hMSC products. 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 in vitro, as many groups are currently promising.

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 recent review article 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.

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!