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

Research output: Contribution to journalArticle

Authors

  • Christopher J. Hewitt
  • Thomas R.j. Heathman
  • Veronica A.m. Glyn
  • Gonҫalo N. Fonte
  • Mariana P. Hanga
  • Karen Coopman
  • Qasim A. Rafiq

Colleges, School and Institutes

Abstract

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 ambr™ 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.

Details

Original languageEnglish
JournalBiochemical Engineering Journal
Early online date6 Aug 2015
Publication statusE-pub ahead of print - 6 Aug 2015

Keywords

  • Human mesenchymal stem cells, Bioreactors, Microcarriers, Mixing, Downstream processing, Harvest