Abstract
ABSTRACTHuman mesenchymal stem cell (hMSC) therapies are currently progressing through clinical development, driving the need for consistent, and cost effective manufacturing processes to meet the lot‐sizes required for commercial production. The use of animal‐derived serum is common in hMSC culture but has many drawbacks such as limited supply, lot‐to‐lot variability, increased regulatory burden, possibility of pathogen transmission, and reduced scope for process optimization. These constraints may impact the development of a consistent large‐scale process and therefore must be addressed. The aim of this work was therefore to run a pilot study in the systematic development of serum‐free hMSC manufacturing process. Human bone‐marrow derived hMSCs were expanded on fibronectin‐coated, non‐porous plastic microcarriers in 100 mL stirred spinner flasks at a density of 3 × 105 cells.mL−1 in serum‐free medium. The hMSCs were successfully harvested by our recently‐developed technique using animal‐free enzymatic cell detachment accompanied by agitation followed by filtration to separate the hMSCs from microcarriers, with a post‐harvest viability of 99.63 ± 0.03%. The hMSCs were found to be in accordance with the ISCT characterization criteria and maintained hMSC outgrowth and colony‐forming potential. The hMSCs were held in suspension post‐harvest to simulate a typical pooling time for a scaled expansion process and cryopreserved in a serum‐free vehicle solution using a controlled‐rate freezing process. Post‐thaw viability was 75.8 ± 1.4% with a similar 3 h attachment efficiency also observed, indicating successful hMSC recovery, and attachment. This approach therefore demonstrates that once an hMSC line and appropriate medium have been selected for production, multiple unit operations can be integrated to generate an animal component‐free hMSC production process from expansion through to cryopreservation. Biotechnol. Bioeng. 2015;112: 1696–1707. © 2015 The Authors. Biotechnology and Bioengineering Published by Wiley Periodicals, Inc.
Highlights
Regenerative medicine (RM) is a growing field that aims to treat currently unmet clinical indications such as diabetes, cardiovascular disease, and neurological disorders by restoring or maintaining tissue function
This study demonstrates for the first time that human Human mesenchymal stem cell (hMSC) can be expanded, harvested, separated, cryopreserved, and recovered from a potentially scalable serum-free microcarrier process
Process mapping is a key part of systematic process development and allows for a structured development methodology centered on the concept of integrated unit operations and is being adopted in current biopharmaceutical manufacture (ICH, 2005)
Summary
Regenerative medicine (RM) is a growing field that aims to treat currently unmet clinical indications such as diabetes, cardiovascular disease, and neurological disorders by restoring or maintaining tissue function. Ever since the term mesenchymal stem cell was first introduced (Caplan, 1991), much anticipation has been generated around the potential for hMSCs to treat and in some cases cure human disease This anticipation has been largely driven by their relative ease of isolation, their ability to proliferate ex vivo under appropriate culture conditions and their capacity to secrete a range of trophic factors which regulate host immune response and initiate tissue repair (Caplan and Dennis, 2006). HMSCs are advancing through clinical development targeting clinical indications such as acute coronary syndrome, stoke, and graft vs host disease (Heathman et al, 2014) Despite this progress, many challenges remain before cost effective production, storage, and delivery of hMSCs to the clinic is feasible.
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