Systematic microcarrier screening and agitated culture conditions improves human mesenchymal stem cell yield in bioreactors

Qasim A. Rafiq, Karen Coopman, Alvin W. Nienow, Christopher J. Hewitt

Research output: Contribution to journalArticle

Abstract

Production of human mesenchymal stem cells for allogeneic cell therapies requires scalable, cost-effective manufacturing processes. Microcarriers enable the culture of anchorage-dependent cells in stirred-tank bioreactors. However, no robust, transferable methodology for microcarrier selection exists, with studies providing little or no reason explaining why a microcarrier was employed. We systematically evaluated 13 microcarriers for human bone marrow-derived MSC (hBM-MSCs) expansion from three donors to establish a reproducible and transferable methodology for microcarrier selection. Monolayer studies demonstrated input cell line variability with respect to growth kinetics and metabolite flux. HBM-MSC1 underwent more cumulative population doublings over three passages in comparison to hBM-MSC2 and hBM-MSC3. In 100 mL spinner flasks, agitated conditions were significantly better than static conditions, irrespective of donor, and relative microcarrier performance was identical where the same microcarriers outperformed others with respect to growth kinetics and metabolite flux. Relative growth kinetics between donor cells on the microcarriers were the same as the monolayer study. Plastic microcarriers were selected as the optimal microcarrier for hBM-MSC expansion. HBM-MSCs were successfully harvested and characterised, demonstrating hBM-MSC immunophenotype and differentiation capacity. This approach provides a systematic method for microcarrier selection, and the findings identify potentially significant bioprocessing implications for microcarrier-based allogeneic cell therapy manufacture. Large-scale production of human bone-marrow derived mesenchymal stem cells (hBM-MSCs) requires expansion on microcarriers in agitated systems. This study demonstrates the importance of microcarrier selection and presents a systematic methodology for selection of an optimal microcarrier. The study also highlights the impact of an agitated culture environment in comparison to a static system, resulting in a significantly higher hBM-MSC yield under agitated conditions.

Original languageEnglish
Pages (from-to)473–486
Number of pages14
JournalBiotechnology Journal
Volume11
Issue number4
Early online date29 Feb 2016
DOIs
Publication statusPublished - Apr 2016

Fingerprint

Bioreactors
Mesenchymal Stromal Cells
Bone Marrow
Cell- and Tissue-Based Therapy
Growth
Plastics
Costs and Cost Analysis
Cell Line
Population

Bibliographical note

© 2015 The Authors. Biotechnology Journal published by WILEY-VCH Verlag GmbH & Co. KGaA, Weinheim. This is an open access article under the terms of the Creative Commons Attribution License, which permits use, distribution and reproduction in any medium, provided the original work is properly cited.

Funding: EPSRC (EP/L015072/1) and Lonza GmbH (Cologne, DE)

Keywords

  • bioreactor
  • cell therapy bioprocessing
  • human mesenchymal stem cell
  • microcarrier
  • regenerative medicine

Cite this

Rafiq, Qasim A. ; Coopman, Karen ; Nienow, Alvin W. ; Hewitt, Christopher J. / Systematic microcarrier screening and agitated culture conditions improves human mesenchymal stem cell yield in bioreactors. In: Biotechnology Journal. 2016 ; Vol. 11, No. 4. pp. 473–486.
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Systematic microcarrier screening and agitated culture conditions improves human mesenchymal stem cell yield in bioreactors. / Rafiq, Qasim A.; Coopman, Karen; Nienow, Alvin W.; Hewitt, Christopher J.

In: Biotechnology Journal, Vol. 11, No. 4, 04.2016, p. 473–486.

Research output: Contribution to journalArticle

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