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

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posted on 2015-10-02, 13:38 authored by Alvin W. Nienow, Christopher J. Hewitt, Thomas Heathman, Veronica A.M. Glyn, G. Fonte, Petra Hanga, Karen CoopmanKaren Coopman, Qasim A. Rafiq
In our recent work in different bioreactors up to 2.5 L in scale, we have successfully cultured hMSCs using the minimum agitator speed required for complete microcarrier suspension, NJS. 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 NJS and detachment in-situ in 15 mL ambrTM bioreactors, 100 mL spinner flasks and 250 mL 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.

Funding

The authors would like to acknowledge the Engineering and Physical Sciences Research Council (EPSRC), Lonza Cologne AG and FUJIFILM Diosynth Biotechnologies for funding this work.

History

School

  • Aeronautical, Automotive, Chemical and Materials Engineering

Department

  • Chemical Engineering

Published in

Biochemical Engineering Journal

Citation

NIENOW, A.W. ...et al., 2016. Agitation conditions for the culture and detachment of hMSCs from microcarriers in multiple bioreactor platforms. Biochemical Engineering Journal, 108, pp. 24–29.

Publisher

© The Authors. Published by Elsevier B.V.

Version

  • VoR (Version of Record)

Publisher statement

This work is made available according to the conditions of the Creative Commons Attribution 4.0 International (CC BY 4.0) licence. Full details of this licence are available at: http://creativecommons.org/licenses/by/4.0/

Publication date

2016

Notes

This is an open access article published by Elsevier under the CC BY license (http://creativecommons.org/licenses/by/4.0/).

ISSN

1873-295X

Language

  • en

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