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

Alvin W. Nienow, Christopher J. Hewitt*, Thomas R.J. Heathman, Veronica A.M. Glyn, Gonsalo N. Fonte, Mariana P. Hanga, Karen Coopman, Qasim A. Rafiq

*Corresponding author for this work

Research output: Contribution to journalArticle

Abstract

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.

Original languageEnglish
Pages (from-to)24-29
Number of pages6
JournalBiochemical Engineering Journal
Volume108
Early online date6 Aug 2015
DOIs
Publication statusPublished - 15 Apr 2016

Bibliographical note

© 2015 The Authors. Published by Elsevier B.V. This is an open access article under the CC BY license(http://creativecommons.org/licenses/by/4.0/).

Funding: (EPSRC), Lonza Cologne AG and FUJIFILM Diosynth Biotechnologies.

Keywords

  • bioreactors
  • downstream processing
  • harvest
  • human mesenchymal stem cells
  • microcarriers
  • mixing

Fingerprint Dive into the research topics of 'Agitation conditions for the culture and detachment of hMSCs from microcarriers in multiple bioreactor platforms'. Together they form a unique fingerprint.

  • Profiles

    No photo of Petra Hanga

    Petra Hanga

    Person: Academic

    Cite this

    Nienow, A. W., Hewitt, C. J., Heathman, T. R. J., Glyn, V. A. M., Fonte, G. N., Hanga, M. P., Coopman, K., & Rafiq, Q. A. (2016). Agitation conditions for the culture and detachment of hMSCs from microcarriers in multiple bioreactor platforms. Biochemical Engineering Journal, 108, 24-29. https://doi.org/10.1016/j.bej.2015.08.003