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.

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:

Towards a Cell Therapy Manufacturing Technology Roadmap -- Resources

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.  

Recently, RoosterBio was part of the first-ever Cell Manufacturing Consortium 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 establish 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.  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.

Relevant Publications:

• Developing Cell Therapy Biomanufacturing Processes
• Cell Therapy Bioprocessing
• Meeting Lot-Size Challenges of Manufacturing Adherent Cells for Therapy
• Developing assays to address identity, potency, purity and safety: cell characterization in cell therapy process development.

NIST Workshop Aims to Educate on Improving Confidence in Measurements Critical for Cell Therapy Products

We are always looking for ways to enable the commercialization of Cell-based Therapies and Technologies.  The ultimate success of the field is dependent on the convergence of several technology fields, and one that isn't given sufficient attention is the Measurement Sciences (or metrology).  The National Institute of Standards and Technology (NIST) has a Biosystems and Biomaterials Division that has several projects focused on their mission of "Building Confidence in Biological Measurements", and they have several people that are working to advance measurements in regenerative medicine.

The NIST BioSystems and Biomaterials Division has several projects related to stem cells and regenerative medicine.

On May 11&12, NIST will be holding a workshop focused on Measurement Assurance for Cell Therapy Products.  The Registration for the workshop is open and is limited to the first 100 registrants, and the Agenda (below) has speakers from Industry, the FDA, NIST, as well as several breakout workshops to focus on some of the analytics that are the low hanging fruit to bring standardization too.

The adage of "Measure Twice, Cut Once" only holds true if you have robust and precise methods and assays, and solid reference standards for which everyone can compare.  Our previous posts on Regenerative Medicine Standardization, and of course the great content on this at the Stem Cell Assays blog are good places for background reading for those interested.

An Open Letter to the Builders of the Cell-based BioEconomy

Dear Stem Cell Pioneer:

February marks the one year anniversary of RoosterBio shipping our first stem cell products to our valued customers, and I personally am very excited for the coming year ahead. We look forward to delivering even more high quality stem cells to people like you that are doing amazing things.

Looking forward into 2015 and beyond, I want to make sure we are staying true to our mission: to greatly increase the availability and accessibility of stem cell technology to researchers and product developers across the globe – and that we are committed to our vision of accelerating the pace of product development in the cellular therapy, bioprinting and tissue engineering markets.  I am hoping to focus our efforts to making sure that we are moving your discoveries and developments forward faster than anticipated.  This will not just be a win for our customers and RoosterBio, but for the entire Regenerative Medicine field.

In 2014 after launching our hMSCs in the unprecedented product format of 10 million cell vials to glowing reviews, we quickly implemented our Starter Kits and Working Cell Bank formats based on your feedback.  These new formats allow for accelerated testing, performance verification, and standardized small scale experimentation with reproducible outcomes.  We also initiated multiple collaborations with leaders in the tissue engineering and biofabrication fields – which has led to multiple conference posters, presentations, and soon to be submitted publications.

We also find ourselves at the precipice of a boom in biofabrication technologies, and we consider this the beginning of the Golden Age of Tissue Engineering.  I anticipate great progress will be made at an increasingly rapid pace.  Now that many of the tools required for bioprinting are becoming “democratized” (simpler, less expensive, more accessible) such as 3D BioPrinters, biomaterials, and primary cells – laboratories can get up and running in a matter of weeks with limited initial resources, something that would have taken months to years and extensive capital in the past.  We are at a special time, and the entire field will be accelerating forward at a rapid pace, making biofabrication truly an exponential medical technology.

2015 will truly be an exciting year for RoosterBio.  We will be participating in a Stem Cell Manufacturing Training Program, helping to organize several conferences on Cell Therapy BioProcessing and BioPrinting, exhibiting at multiple conferences, as well are contributing to initiatives such as the Georgia Tech Cell Manufacturing Consortium and the NIST Workshop on Strategies to Achieve Measurement Assurance for Cell Therapy Products.  The primary motivation behind these initiatives are to make sure that we are driving forward our vision and delivering on our mission.

None of this would be possible without the hard work and dedication of the entire RoosterBio team, 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 BioEconomy.

All the best from Frederick, Maryland.


Jon A Rowley
Chief Executive & Technology Officer
RoosterBio Inc.

Welcome to the Golden Age of BioPrinting, Tissue Engineering and BioFabrication

A"Golden Age" is defined as a period of time in a field where "great tasks are accomplished."  The ancient Greek philosopher Hesiod initially coined this phrase, and I think if he were alive today, he would agree with us that we are in a special time of technology convergence where innovations and advancements are progressing at an accelerating rate.  The fields of Tissue Engineering and Regenerative Medicine are benefiting from these rapid technology advancements.



We are now at the beginnings of the Golden Age of BioFabrication.  The last 20 years has seen steady progress in the Tissue Engineering field, but the cost and time it has taken to develop products based on these technologies has been prohibitive.  Thus, only the best funded labs have been able to perform this very expensive R&D.  Within the last year, products such as high volume stem cells (via RoosterBio) and low cost bioprinters (from our collaborators BioBots) have been coming to market and dramatically reduce the cost, the time, and the complexity to fabricate three dimensional biological structures that are the precursors to tomorrow's tissue engineered products.  By removing the technology and cost barriers and democratizing biofabrication technology, more labs can now afford to do the applied R&D, allowing more work to be accomplished faster, completely changing the equation of how labs function.  This is accelerating the development of this entire field.

 Walter Isaacson makes the point over and over in his new book The Innovators that collaboration between people and groups with complementary skill sets is essential to innovation and technology progress.  We, at RoosterBio, have always said that communication platforms (such as social networks, conferences, biohacker spacers, blogs, and journals) are also critical for those in a field to share knowledge and experiences - further progressing the thought convergence.  This February 9th and 10th in Boston is a focused conference on Tissue Engineering and BioPrinting that SelectBio is hosting.  The top researchers, thought leaders, and product developers in the field will be presenting cutting edge research, technology development, and commercialization strategies.  We hope to see you there.

Rapid and Economic Generation of hMSC Spheroids for Macroscopic Tissue Biofabrication

Mesenchymal stem cells (MSCs) aggregated into three-dimensional (3D) cellular spheroids are a potent configuration for cell therapy and tissue engineering research and product development (5), and cellular spheroids are a preferred format for many bioprinting applications (10).  Cellular spheroids are essentially micro-tissues that can be manufactured as standardized “living materials” with certain controllable, measurable, and evolving material properties (10). Studies have shown that MSC aggregation into spheroids yield improved in vitro biological functionality over MSCs grown as 2D monolayer; likely due to the 3D tissue-like structure resembling the native configuration of cells in vivo with a microenvironment that allows for direct cell-cell signaling and cell-matrix interactions. MSC spheroids demonstrate enhanced cartilage, bone, and fat differentiation, as well as increased paracrine factor secretion over 2D MSC cultures (1-7). In vivo administration of hMSC spheroids has also showed enhanced therapeutic properties in pre-clinical models of myocardial infarction, bone and cartilage repair, and limb ischemia (3, 5, 8).  

Traditionally, aggregates were formed using suspension culture in spinners or shake flasks or in hanging drop cultures (5). The advancement of technologies has allowed one to quickly and easily generate large numbers of spheroids consistent in size and shape using forced aggregation in micro-wells (AggreWells, Stem Cell Technologies), or using liquid handling automation and 96 or 384 well hanging-droplet plates. There are also tools available that allow researchers to mold micro-tissues into interesting shapes such as rods, toroids, honeycombs, or whatever one can dream up (12-13).  While there are multiple methods for creating hMSC micro-tissues, the biggest challenge is reproducibly growing up sufficient hMSCs to create enough spheroids to start an experiment.  For example, if a researcher needs 10,000 spheroids with an average of 1000 cells per spheroid, then he/she will need at least 10 million cells to begin the experiment, which can take weeks to grow (see process flow diagrams below).  If he/she wishes to use 5000, or 10,000 cells per spheroid, then he/she will need 50 million or 100 million cells for his/her experiment.  This volume of cells has traditionally been very costly and time consuming to generate.

This Application Blog Post will provide a simple protocol to rapidly and economically generate tens of millions of high quality hMSCs so that researchers can minimize their time spent on routine cell culture and maximize their effort on performing hMSC spheroid-based experiments.

How Quickly are Cell-based Products Really Developing? Thoughts from the IBC Cell Therapy Bioprocessing Conference

Last week was the 4^th Annual IBC Cell Therapy Bioprocessing Conference.  IBC (home of the BioProcess International conference) was the first conference organizer to dedicate a focused meeting on Cell Therapy Manufacturing Technologies 4 years ago.  Since the first conference in 2011, the growth in the field, and the conference, has been amazing.  The attendance has grown from less than 90 in year 1 to over 200 this year.  The content has also evolved heavily over the last 4 years, demonstrating a high level of sophistication and maturity in a field that seems “early stage” to those looking in from the outside.  The talks this year increased in the amount and quality of data presented. Topics included the impact of automation on the simplification, streamlining, and cost reduction of autologous therapies, the use of Quality by Design (QbD) in bioreactor scale-up and analytical development, advances in tissue engineering and biofabrication techniques, and even 2 year data on marketed products.   Phil Vanek, the General Manager of GE Healthcare’s Cell Therapy business, summed it up during his talk where he stated that: GE is interested in 1) big problems, 2) compelling clinical data, and 3) opportunities for “industrialization”, and “Cell Therapy/Regenerative Medicine has all three”.

Various cell manufacturing and processing devices seen throughout the exhibits at IBC's
4th Annual Cell Therapy BioProcessing Conference - No BioPrinters (yet!)

There are many signs that the Cell Therapy field is moving much faster than the protein therapeutics field before it and demonstrating rapid progress.  What we have here is a traditional case of  “standing on the shoulders of giants”, which has been paraphrased on Wikipedia as "discovering truth by building on previous discoveries”.  

Welcome to the RoosterBio Blog

Everyone at RoosterBio is extremely excited to be launching our company, shipping our first products, and interacting with tissue engineers, cell therapists, synthetic biologists, and our customers to learn about innovative research in these fields, share our knowledge, and contribute to the Cell and Tissue Engineering Revolution. The central theme behind RoosterBio, or what we call our “business hypothesis”, is that as living cellular technologies become more affordable, easier to access, and much simpler to incorporate into product development efforts– there will be a rapid acceleration in products coming to market that incorporate these technologies. We believe that we can help shape this new market by Democratizing Cell Technologies and making them abundant, affordable, and much simpler to translate into the clinic.

We have assembled a team at RoosterBio that have years of experience in stem cell R&D, cell therapeutic product and process development, and manufacturing operations, and with this blog we hope to share many of the topics that we are so enthusiastic about.  We gravitate toward technologies at the technical interfaces of fields such as 3D printing and tissue engineering; thus 3D bioprinting of tissues will be a big topic on this blog.  We will be sharing our thoughts on step-changes in technology on topics like stem cell therapies, engineered tissues and organs, biological robots, manufacturing technologies, and synthetic biology.  We will also engage in educational posts related to our expertise, and comment on broad themes that are facing the industries that we care about, such as establishing standards in the stem cell arena.  The developments in progressive fields always have profound effects on science and society, and we believe we are on the verge of the cell and tissue engineering revolution. 

This blog is targeted at the scientists, technologists, engineers, and doctors that are working passionately and feverishly to bring cell-based products to patients, as well as for general audiences that are interested in the driving forces behind cellular therapies, regenerative medicines, and tissue engineering technologies.  We hope you will come back to learn and engage in the conversation, and also be a part of the next technology revolution as Biology becomes Technology. 

We encourage everyone to leave comments, and to feel free to say what is on your mind. We look forward to the dialogue and helping to accelerate the Cell-based BioEconomy!

- The RoosterBio Team