September 18, 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

Authored by Iain Farrance, PhD, Technical Marketing Associate


Introduction

Human mesenchymal stromal cells (hMSC) are considered the "workhorse" of Regenerative Medicine (RegenMed).  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 (clinical trials.gov).  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. Our goal is to have the same impact on the RegenMed industry that Intel had on the computer industry.

Use of 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.  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 Tissue RegenerationTherapeutics Inc. (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.



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). RoosterBio’s launch of our XF hUC-MSC (RoosterVial™-hUC-MSC-XF) 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 here.

RoosterBio’s RoosterVial-hUC-MSC-XF and RoosterNourish™-MSC cell and medium bioprocess system 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 is the only hUC-MSC commercially available with a clear line of sight to clinical translation.  Additionally, 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.  Finally, RoosterBio provides first in class characterization of hMSC key quality attributes (PDL, identity, expansion potential) and functional assays (cytokine secretion, trilineage differentiation, immunomodulation).

August 16, 2019

Behind Our Best Places to Work Award: Life Inside The Roost


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. 

What Makes RoosterBio a “Best Place to Work”

The Frederick County Best Places to Work 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!

Take a peek inside life at The Roost.


Attracting the Best Talent

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.

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. 

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.

Retaining a Strong Workforce

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.

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.


Cool Perks

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.

Promoting Fun at Work

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 Make Work Awesome. 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.


Celebrating Success Together

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.

Not Just Culture Cluck

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 LinkedIn and Facebook for testimonials from almost half of the company on what they decided to join RoosterBio. 

Check out our open positions as we expand our presence across the globe and be a part of this exciting journey.

July 15, 2019

Advances in Clinical Translation and Scale-up of MSCs and Extracellular Vesicles at ISCT 2019

Authored by Katrina Adlerz, Ph.D., Scientist, Analytics, Product & Process Development
& Josephine Lembong, Ph.D., Scientist, Analytics, Product & Process Development

The 2019 International Society for Cell and Gene Therapy (ISCT) Annual Meeting 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 Global Clinical Trial Landscape of MSCs 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 Tissue Regeneration Therapeutics to make umbilical cord -derived MSCs broadly available. 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.

MSC Progress in Clinical Trials, Manufacturing, and Comparability

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 Phase III clinical trials for ischemic stroke patients 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 Alofisel, an allogeneic MSC product that has been approved in Europe for perianal fistulas in Crohn’s disease. Some challenges in clinical trials were highlight
Photo courtesy http://www.isct2019.com/photo-gallery/
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 Population Doubling Level (PDL). Furthermore, in his recent study where fresh and cryopreserved cells were

June 11, 2019

Cells as Bioinks for 3D Bioprinting

Authored by Mayasari Lim, PhD, Regional Account Manager, West Coast, RoosterBio

Bioprinting overview

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.

Which cell should I use?

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.

How many cells do I need?

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.


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 (RoosterRTP™) 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 previous blog 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.

References:

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

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



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:
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 average of 3 to 4 years. At the same time, it is critical for researchers and product developers to shorten both publication and product development timeframes. RoosterBio high volume hMSCs and bioprocess media systems 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.

Highlighted Publications:

Defining Hydrogel Properties to Instruct Lineage- and Cell-Specific Mesenchymal Differentiation. Hung BP, Harvestine JN, Saiz AM, Gonzalez-Fernandez T, Sahar DE, Weiss ML, Leach JK, Biomaterials, 2019.

IFN-γ and TNF-α Pre-licensing Protects Mesenchymal Stromal Cells from the Pro-inflammatory Effects of Palmitate. Boland L, Burand AJ, Brown AJ, Boyt D, Lira VA, Ankrum JA. Mol Ther. 2018.

Deciphering the role of substrate stiffness in enhancing the internalization efficiency of plasmid DNA in stem cells using lipid-based nanocarriers. Modaresi S, Pacelli S, Whitlow J, Paul A. Nanoscale. 2018.

Acoustophoretic printing. Foresti D, Kroll KT, Amissah R, Sillani F, Homan KA, Poulikakos D, Lewis JA. Sci Adv. 2018.

3D printed biofunctionalized scaffolds for microfracture repair of cartilage defects. Guo T, Noshin M, Baker HB, Taskoy E, Meredith SJ, Tang Q, Ringel JP, Lerman MJ, Chen Y, Packer JD, Fisher JP. Biomaterials. 2018.

Mesenchymal stem cell-derived extracellular vesicles attenuate pulmonary vascular permeability and lung injury induced by hemorrhagic shock and trauma. 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 Trauma Acute Care Surg. 2018.

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.


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.

References:
  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.