Serum-free process development:improving the yield and consistency of human mesenchymal stromal cell production

Heathman, Thomas R.J., Stolzing, Alexandra, Fabian, Claire, Rafiq, Qasim A., Coopman, Karen, Nienow, Alvin W., Kara, Bo and Hewitt, Christopher J. (2015). Serum-free process development:improving the yield and consistency of human mesenchymal stromal cell production. Cytotherapy, 17 (11), pp. 1524-1535.

Abstract

Background aims: The cost-effective production of human mesenchymal stromal cells (hMSCs) for off-the-shelf and patient specific therapies will require an increasing focus on improving product yield and driving manufacturing consistency. Methods: Bone marrow-derived hMSCs (BM-hMSCs) from two donors were expanded for 36 days in monolayer with medium supplemented with either fetal bovine serum (FBS) or PRIME-XV serum-free medium (SFM). Cells were assessed throughout culture for proliferation, mean cell diameter, colony-forming potential, osteogenic potential, gene expression and metabolites. Results: Expansion of BM-hMSCs in PRIME-XV SFM resulted in a significantly higher growth rate (P < 0.001) and increased consistency between donors compared with FBS-based culture. FBS-based culture showed an inter-batch production range of 0.9 and 5 days per dose compared with 0.5 and 0.6 days in SFM for each BM-hMSC donor line. The consistency between donors was also improved by the use of PRIME-XV SFM, with a production range of 0.9 days compared with 19.4 days in FBS-based culture. Mean cell diameter has also been demonstrated as a process metric for BM-hMSC growth rate and senescence through a correlation (R<sup>2</sup> = 0.8705) across all conditions. PRIME-XV SFM has also shown increased consistency in BM-hMSC characteristics such as per cell metabolite utilization, in vitro colony-forming potential and osteogenic potential despite the higher number of population doublings. Conclusions: We have increased the yield and consistency of BM-hMSC expansion between donors, demonstrating a level of control over the product, which has the potential to increase the cost-effectiveness and reduce the risk in these manufacturing processes.

Publication DOI: https://doi.org/10.1016/j.jcyt.2015.08.002
Divisions: Life & Health Sciences
Additional Information: © 2015, International Society for Cellular Therapy. Published by Elsevier Inc. This is an open access article under the CC BY license (http://creativecommons.org/licenses/by/4.0/). Funding: EPSRC; and FUJIFILM Diosynth Biotechnologies. Supplementary data related to this article can be found at http://dx.doi.org/10.1016/j.jcyt.2015.08.002
Uncontrolled Keywords: cell-based therapy,comparability,consistency,human mesenchymal stromal cell,manufacturing,regenerative medicine,serum-free,yield,Immunology,Immunology and Allergy,Oncology,Genetics(clinical),Transplantation,Cancer Research,Cell Biology
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Related URLs: http://www.scop ... tnerID=8YFLogxK (Scopus URL)
Published Date: 2015-11
Authors: Heathman, Thomas R.J.
Stolzing, Alexandra
Fabian, Claire
Rafiq, Qasim A.
Coopman, Karen
Nienow, Alvin W.
Kara, Bo
Hewitt, Christopher J. ( 0000-0001-6988-6071)

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