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|Title: ||Expansion, harvest and cryopreservation of human mesenchymal stem cells in a serum-free microcarrier process|
|Authors: ||Heathman, Thomas R.J.|
Glyn, Veronica A.M.
Rafiq, Qasim A.
Nienow, Alvin W.
Hewitt, Christopher J.
Human mesenchymal stem cell
|Issue Date: ||2015|
|Publisher: ||Wiley / © The Authors|
|Citation: ||HEATHMAN, T.R.J. ... et al, 2015. Expansion, harvest and cryopreservation of human mesenchymal stem cells in a serum-free microcarrier process. Biotechnology and Bioengineering, 112 (8), pp.1696–1707|
|Abstract: ||Human mesenchymal stem cell (hMSC) therapies are currently progressing through clinical development, driving the need for consistent, and cost effective manufacturing processes to meet the lot-sizes required for commercial production. The use of animal-derived serum is common in hMSC culture but has many drawbacks such as limited supply, lot-to-lot variability, increased regulatory burden, possibility of pathogen transmission, and reduced scope for process optimization. These constraints may impact the development of a consistent large-scale process and therefore must be addressed. The aim of this work was therefore to run a pilot study in the systematic development of serum-free hMSC manufacturing process. Human bone-marrow derived hMSCs were expanded on fibronectin-coated, non-porous plastic microcarriers in 100 mL stirred spinner flasks at a density of 3 × 105 cells.mL−1 in serum-free medium. The hMSCs were successfully harvested by our recently-developed technique using animal-free enzymatic cell detachment accompanied by agitation followed by filtration to separate the hMSCs from microcarriers, with a post-harvest viability of 99.63 ± 0.03%. The hMSCs were found to be in accordance with the ISCT characterization criteria and maintained hMSC outgrowth and colony-forming potential. The hMSCs were held in suspension post-harvest to simulate a typical pooling time for a scaled expansion process and cryopreserved in a serum-free vehicle solution using a controlled-rate freezing process. Post-thaw viability was 75.8 ± 1.4% with a similar 3 h attachment efficiency also observed, indicating successful hMSC recovery, and attachment. This approach therefore demonstrates that once an hMSC line and appropriate medium have been selected for production, multiple unit operations can be integrated to generate an animal component-free hMSC production process from expansion through to cryopreservation.|
|Description: ||This is an open-access article published by Wiley and 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/4.0/|
|Sponsor: ||This study has been funded by the Engineering and Physical Sciences
Research Council (EPSRC), the Biotechnology and Biological Sciences
Research Council (BBSRC) and FUJIFILM Diosynth Biotechnologies.|
|Publisher Link: ||http://dx.doi.org/10.1002/bit.25582|
|Appears in Collections:||Published Articles (Chemical Engineering)|
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