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

Authors
  • Rafiq, Qasim A.1
  • Brosnan, Kathryn M.1
  • Coopman, Karen1
  • Nienow, Alvin W.1, 2
  • Hewitt, Christopher J.1
  • 1 Loughborough University, Leicestershire, LE11 3TU, UK , Leicestershire (United Kingdom)
  • 2 University of Birmingham, Edgbaston, Birmingham, B15 2TT, UK , Birmingham (United Kingdom)
Type
Published Article
Journal
Biotechnology Letters
Publisher
Springer-Verlag
Publication Date
Apr 23, 2013
Volume
35
Issue
8
Pages
1233–1245
Identifiers
DOI: 10.1007/s10529-013-1211-9
Source
Springer Nature
Keywords
License
Yellow

Abstract

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.

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