Optimization of agitation speed in spinner flask for microcarrier structural integrity and expansion of induced pluripotent stem cells

In recent times, the study and use of induced pluripotent stem cells (iPSC) have become important in order to avoid the ethical issues surrounding the use of embryonic stem cells. Therapeutic, industrial and research based use of iPSC requires large quantities of cells generated in vitro. Mammalian...

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Bibliographic Details
Published in:Cytotechnology (Dordrecht) 2016-01, Vol.68 (1), p.45-59
Main Authors: Gupta, Priyanka, Ismadi, Mohd-Zulhilmi, Verma, Paul J., Fouras, Andreas, Jadhav, Sameer, Bellare, Jayesh, Hourigan, Kerry
Format: Article
Language:English
Subjects:
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Agitation
Biochemistry
Biomedicine
Biotechnology
Cell culture
Cell survival
Chemistry
Chemistry and Materials Science
Embryo cells
Mammalian cells
Original Research
Pluripotency
Stem cells
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Summary:In recent times, the study and use of induced pluripotent stem cells (iPSC) have become important in order to avoid the ethical issues surrounding the use of embryonic stem cells. Therapeutic, industrial and research based use of iPSC requires large quantities of cells generated in vitro. Mammalian cells, including pluripotent stem cells, have been expanded using 3D culture, however current limitations have not been overcome to allow a uniform, optimized platform for dynamic culture of pluripotent stem cells to be achieved. In the current work, we have expanded mouse iPSC in a spinner flask using Cytodex 3 microcarriers. We have looked at the effect of agitation on the microcarrier survival and optimized an agitation speed that supports bead suspension and iPS cell expansion without any bead breakage. Under the optimized conditions, the mouse iPSC were able to maintain their growth, pluripotency and differentiation capability. We demonstrate that microcarrier survival and iPS cell expansion in a spinner flask are reliant on a very narrow range of spin rates, highlighting the need for precise control of such set ups and the need for improved design of more robust systems.
ISSN:0920-9069
1573-0778
DOI:10.1007/s10616-014-9750-z