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

Thomas R.J. Heathman, Alexandra Stolzing, Claire Fabian, Qasim A. Rafiq, Karen Coopman, Alvin W. Nienow, Bo Kara, Christopher J. Hewitt*

*Corresponding author for this work

Research output: Contribution to journalArticle

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.

Original languageEnglish
Pages (from-to)1524-1535
Number of pages12
JournalCytotherapy
Volume17
Issue number11
Early online date30 Sep 2015
DOIs
Publication statusPublished - Nov 2015

Bibliographical note

© 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

Keywords

  • cell-based therapy
  • comparability
  • consistency
  • human mesenchymal stromal cell
  • manufacturing
  • regenerative medicine
  • serum-free
  • yield

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    Heathman, T. R. J., Stolzing, A., Fabian, C., Rafiq, Q. A., Coopman, K., Nienow, A. W., Kara, B., & Hewitt, C. J. (2015). Serum-free process development: improving the yield and consistency of human mesenchymal stromal cell production. Cytotherapy, 17(11), 1524-1535. https://doi.org/10.1016/j.jcyt.2015.08.002