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Culture of human mesenchymal stem cells on microcarriers in a 5 l stirred-tank bioreactor

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journal contribution
posted on 06.06.2014 by Qasim Rafiq, Kathryn Brosnan, Karen Coopman, Alvin W. Nienow, Christopher Hewitt
For the first time, fully functional human mesenchymal stem cells (hMSCs) have been cultured at the litre-scale on microcarriers in a stirred-tank 5 l bioreactor, (2.5 l working volume) and were harvested via a potentially scalable detachment protocol that allowed for the successful detachment of hMSCs from the cell-microcarrier suspension. Over 12 days, the dissolved O2 concentration was >45 % of saturation and the pH between 7.2 and 6.7 giving a maximum cell density in the 5 l bioreactor of 1.7 × 105 cells/ml; this represents >sixfold expansion of the hMSCs, equivalent to that achievable from 65 fully-confluent T-175 flasks. During this time, the average specific O2 uptake of the cells in the 5 l bioreactor was 8.1 fmol/cell h and, in all cases, the 5 l bioreactors outperformed the equivalent 100 ml spinner-flasks run in parallel with respect to cell yields and growth rates. In addition, yield coefficients, specific growth rates and doubling times were calculated for all systems. Neither the upstream nor downstream bioprocessing unit operations had a discernible effect on cell quality with the harvested cells retaining their immunophenotypic markers, key morphological features and differentiation capacity.

History

School

  • Aeronautical, Automotive, Chemical and Materials Engineering

Department

  • Chemical Engineering

Published in

BIOTECHNOLOGY LETTERS

Volume

35

Issue

8

Pages

1233 - 1245 (13)

Citation

RAFIQ, Q. ... et al., 2013. Culture of human mesenchymal stem cells on microcarriers in a 5 l stirred-tank bioreactor. Biotechnology Letters, 35 (8), pp. 1233 - 1245.

Publisher

© Springer Science+Business Media

Version

AM (Accepted Manuscript)

Publication date

2013

Notes

This article was published in the journal, Biotechnology Letters [© Springer Science+Business Media] and the definitive version is available at: http://dx.doi.org/10.1007/s10529-013-1211-9

ISSN

0141-5492

Language

en

Exports