Optimization of agitation speed in spinner flask for microcarrier structural integrity and expansion of induced pluripotent stem cells
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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.
KeywordsMicrocarrier Induced pluripotent stem cells Spinner flask
The authors would like to acknowledge Ms. Joan Clark, Monash Micro Imaging Facility (Monash University), for helping with the SEM imaging, Dr. Trevor Wilson, Medical Genomics Facility, Monash Health and Technology Precinct, for his help in the Array Scan analysis, and Ms. Karla Contreras, Division of Biological Engineering (Monash University) for her assistance. This work was supported by the Australian Research Council Discovery Program (Grant DPDP130100822) and by the Australia India Strategic Research Fund (Grant BF050038).
Conflict of interest
The authors declare that no competing financial interests exist for this work.
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