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


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.

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Divisions: College of Health & Life Sciences
Additional Information: © 2015, International Society for Cellular Therapy. Published by Elsevier Inc. This is an open access article under the CC BY license ( Funding: EPSRC; and FUJIFILM Diosynth Biotechnologies. Supplementary data related to this article can be found at
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
Publication ISSN: 1477-2566
Last Modified: 10 Apr 2024 07:12
Date Deposited: 22 Oct 2015 14:55
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Related URLs: http://www.scop ... tnerID=8YFLogxK (Scopus URL)
PURE Output Type: Article
Published Date: 2015-11
Published Online Date: 2015-09-30
Authors: Heathman, Thomas R.J.
Stolzing, Alexandra
Fabian, Claire
Rafiq, Qasim A.
Coopman, Karen
Nienow, Alvin W.
Kara, Bo
Hewitt, Christopher J. (ORCID Profile 0000-0001-6988-6071)



Version: Published Version

License: Creative Commons Attribution

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