Agitation conditions for the culture and detachment of hMSCs from microcarriers in multiple bioreactor platforms


In our recent work in different bioreactors up to 2.5L in scale, we have successfully cultured hMSCs using the minimum agitator speed required for complete microcarrier suspension, N <inf>JS.</inf> In addition, we also reported a scaleable protocol for the detachment from microcarriers in spinner flasks of hMSCs from two donors. The essence of the protocol is the use of a short period of intense agitation in the presence of enzymes such that the cells are detached; but once detachment is achieved, the cells are smaller than the Kolmogorov scale of turbulence and hence not damaged. Here, the same approach has been effective for culture at N <inf>JS</inf> and detachment in-situ in 15mL ambr™ bioreactors, 100mL spinner flasks and 250mL Dasgip bioreactors. In these experiments, cells from four different donors were used along with two types of microcarrier with and without surface coatings (two types), four different enzymes and three different growth media (with and without serum), a total of 22 different combinations. In all cases after detachment, the cells were shown to retain their desired quality attributes and were able to proliferate. This agitation strategy with respect to culture and harvest therefore offers a sound basis for a wide range of scales of operation.

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Divisions: College of Health & Life Sciences
College of Health & Life Sciences > School of Biosciences > Cell & Tissue Biomedical Research
Additional Information: © 2015 The Authors. Published by Elsevier B.V. This is an open access article under the CC BY license( Funding: (EPSRC), Lonza Cologne AG and FUJIFILM Diosynth Biotechnologies.
Uncontrolled Keywords: bioreactors,downstream processing,harvest,human mesenchymal stem cells,microcarriers,mixing,Biotechnology,Bioengineering,Biomedical Engineering,Environmental Engineering
Publication ISSN: 1873-295X
Last Modified: 08 Jul 2024 07:19
Date Deposited: 24 Sep 2015 11:40
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Related URLs: http://www.scop ... tnerID=8YFLogxK (Scopus URL)
PURE Output Type: Article
Published Date: 2016-04-15
Published Online Date: 2015-08-06
Accepted Date: 2015
Authors: Nienow, Alvin W.
Hewitt, Christopher J. (ORCID Profile 0000-0001-6988-6071)
Heathman, Thomas R.J.
Glyn, Veronica A.M.
Fonte, Gonsalo N.
Hanga, Mariana P. (ORCID Profile 0000-0002-2427-4052)
Coopman, Karen
Rafiq, Qasim A.



Version: Published Version

License: Creative Commons Attribution

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