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Title: A potentially scalable method for the harvesting of hMSCs from microcarriers
Authors: Nienow, Alvin W.
Rafiq, Qasim A.
Coopman, Karen
Hewitt, Christopher J.
Keywords: Human mesenchymal stem cells
Microcarriers
Large scale processing
Downstream processing
Cell harvest
Agitation
Issue Date: 2014
Publisher: Elsevier B.V. / © The Authors
Citation: NIENOW, A.W. ... et al, 2014. A potentially scalable method for the harvesting of hMSCs from microcarriers. Biochemical Engineering Journal, 85, pp. 79 - 88.
Abstract: The use of hMSCs for allogeneic therapies requiring lot sizes of billions of cells will necessitate large-scale culture techniques such as the expansion of cells on microcarriers in bioreactors. Whilst much research investigating hMSC culture on microcarriers has focused on growth, much less involves their harvesting for passaging or as a step towards cryopreservation and storage. A successful new harvesting method has recently been outlined for cells grown on SoloHill microcarriers in a 5 L bioreactor [1]. Here, this new method is set out in detail, harvesting being defined as a two-step process involving cell ‘detachment’ from the microcarriers’ surface followed by the ‘separation’ of the two entities. The new detachment method is based on theoretical concepts originally developed for secondary nucleation due to agitation. Based on this theory, it is suggested that a short period (here 7 min) of intense agitation in the presence of a suitable enzyme should detach the cells from the relatively large microcarriers. In addition, once detached, the cells should not be damaged because they are smaller than the Kolmogorov microscale. Detachment was then successfully achieved for hMSCs from two different donors using microcarrier/cell suspensions up to 100 mL in a spinner flask. In both cases, harvesting was completed by separating cells from microcarriers using a Steriflip® vacuum filter. The overall harvesting efficiency was >95% and after harvesting, the cells maintained all the attributes expected of hMSC cells. The underlying theoretical concepts suggest that the method is scalable and this aspect is discussed too.
Description: This is an open-access article distributed under the terms of the Creative Commons Attribution License, which permits unrestricted use, distribution, and reproduction in any medium, provided the original author and source are credited. Full details of the CC BY licence are available at: http://creativecommons.org/licenses/by/3.0/
Sponsor: The authors would like to acknowledge the Biotechnology and Biological Sciences Research Council (BBSRC; UK) Bioprocessing Research Industries Club (BRIC), the Engineering and Physical Sciences Research Council (EPSRC; UK) [grant number EP/F500491/1] and Lonza GmbH (Cologne, Germany) for their support and funding.
Version: Published
DOI: 10.1016/j.bej.2014.02.005
URI: https://dspace.lboro.ac.uk/2134/14883
Publisher Link: http://dx.doi.org/10.1016/j.bej.2014.02.005
ISSN: 1369-703X
Appears in Collections:Published Articles (Chemical Engineering)

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