Abstract
Efficient cryopreservation of human pluripotent stem cells (hPSCs) in chemically defined, xeno-free conditions is highly desirable for medical research and clinical applications such as cell-based therapies. Here we present a simple and effective slow freezing–rapid thawing protocol for the cryopreservation of feeder-free, single hPSCs. This cryopreservation protocol involves the supplementation of 10 % dimethyl sulfoxide (DMSO) and 10 μM Rho-associated kinase inhibitor Y-27632 into two types of xeno-free, defined media supplements (Knockout Serum Replacement and TeSR2). High post-thaw cell recovery (~90 %) and cell expansion (~70 %) can be achieved using this protocol. The cryopreserved single cells retain the morphological characteristics of hPSCs and differentiation capabilities of pluripotent stem cells.
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Acknowledgements
This work was supported by a CIHR Regenerative Medicine Team Grant FRN 27639.
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Meng, G., Poon, A., Liu, S., Rancourt, D.E. (2016). An Effective and Reliable Xeno-free Cryopreservation Protocol for Single Human Pluripotent Stem Cells. In: Turksen, K. (eds) Stem Cell Heterogeneity. Methods in Molecular Biology, vol 1516. Humana Press, New York, NY. https://doi.org/10.1007/7651_2016_322
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DOI: https://doi.org/10.1007/7651_2016_322
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