RESEARCH ARTICLE


Expansion and Harvesting of hMSC-TERT



Christian Weber1, Sebastian Pohl1, Ralf Pörtner2, Christine Wallrapp3, Moustapha Kassem4, Peter Geigle3, Peter Czermak*, 1, 5
1 Institute of Biopharmaceutical Technology, University of Applied Sciences Giessen-Friedberg, Giessen-Germany
2 Institute of Bioprocess and Biosystems Engineering, University of Technology, Hamburg-Germany
3 CellMed AG, Alzenau-Germany
4 Department of Endocrinology and Metabolism, University Hospital of Odense, Odense-Denmark5
5 Department of Chemical Engineering, Kansas State University, Manhattan KS-USA


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2007 Bentham Science Publishers Ltd.

open-access license: This is an open access article distributed under the terms of the Creative Commons Attribution License (http://creativecommons.org/licenses/by/2.5/) which permits unrestrictive use, distribution, and reproduction in any medium, provided the original work is properly cited.

* Address correspondence to this author at the Dipl.-Biol. Dipl.-Ing. Christian Weber, Institute of Biopharmaceutical Technology, University of Applied Sciences Giessen-Friedberg, Wiesenstrasse 14, 35390 Giessen, Germany; E-mail: Peter.Czermak@tg.fh-giessen.de


Abstract

The expansion of human mesenchymal stem cells as suspension culture by means of spinner flasks and microcarriers, compared to the cultivation in tissue culture flasks, offers the advantage of reducing the requirements of large incubator capacities as well as reducing the handling effort during cultivation and harvesting. Nonporous microcarriers are preferable when the cells need to be kept in viable condition for further applications like tissue engineering or cell therapy. In this study, the qualification of Biosilon, Cytodex 1, Cytodex 3, RapidCell and P102-L for expansion of hMSC-TERT with an associated harvesting process using either trypsin, accutase, collagenase or a trypsin-accutase mixture was investigated. A subsequent adipogenic differentiation of harvested hMSC-TERT was performed in order to observe possible negative effects on their (adipogenic) differentiation potential as a result of the cultivation and harvesting method. The cultivated cells showed an average growth rate of 0.52 d-1. The cells cultivated on Biosilon, RapidCell and P102-L were harvested succesfully achieving high cell yield and vitalities near 100%. This was not the case for cells on Cytodex 1 and Cytodex 3. The trypsin-accutase mix was most effective. After spinner expansion and harvesting the cells were successfully differentiated to adipocytes.

Keywords: Harvest, hMSC-TERT, mesenchymal stem cells, microcarrier, spinner flask..