Expansion and Harvesting of hMSC-TERT

Abstract

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.

Process control in cell culture technology using dielectric spectroscopy

“…The growing area of regenerative medicine including cell therapy and gene therapy with examples such as production of encapsulated mesenchymal stem cells could be considered a good application for DS. , Controlled cell expansion is often required for mesenchymal stem cell production [Weber et al, 2008, Weber, et al, 2007a, Weber, et al, 2007b. In-line monitoring by DS for cell density and the time for culture harvest might be especially useful when stem cells are grown adherent or on carriers in fixed bed reactors since it is difficult to take samples.…”

“…The production processes for cells or cell related products are complex, and must comply with GMP requirements [Baldi, et al, 2007, Butler, 2005, Weber et al, 2008, Weber et al, 2007a, Weber, et al, 2007b, Wurm, 2004. The intricate relationships of process parameters and process outcome may not be clear in all cases, especially on the industrial scale.…”

Mixing theory for culture and harvest in bioreactors of human mesenchymal stem cells on microcarriers

“…Typically, cell detachment from microcarriers is by enzymatic digestion [2] aided by extensional flow using a pipette. At larger scales, such an approach becomes impractical and in addition, the enzymes can also damage the cells if exposure is prolonged.…”

CELL DETACHMENT BY PROLYL-SPECIFIC ENDOPEPTIDASE FROM <i>WOLFIPORIA COCOS</i>

“…Suitable growth surfaces of these cells are often made of polystyrene. Polystyrene is offered as cell culture plates for R&D or as spherical carriers for bioreactor processes (Weber et al, 2007). A precondition for cell attachment is a protein adsorption to the polystyrene.…”