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Slow Cooling Cryopreservation Optimized to Human Pluripotent Stem Cells

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Biobanking and Cryopreservation of Stem Cells

Part of the book series: Advances in Experimental Medicine and Biology ((AEMB,volume 951))

Abstract

Human pluripotent stem cells (hPSCs) have the potential for unlimited expansion and differentiation into cells that form all three germ layers. Cryopreservation is one of the key processes for successful applications of hPSCs, because it allows semi-permanent preservation of cells and their easy transportation. Most animal cell lines, including mouse embryonic stem cells, are standardly cryopreserved by slow cooling; however, hPSCs have been difficult to preserve and their cell viability has been extremely low whenever cryopreservation has been attempted.

Here, we investigate the reasons for failure of slow cooling in hPSC cryopreservation. Cryopreservation involves a series of steps and is not a straightforward process. Cells may die due to various reasons during cryopreservation. Indeed, hPSCs preserved by traditional methods often suffer necrosis during the freeze-thawing stages, and the colony state of hPSCs prior to cryopreservation is a major factor contributing to cell death.

It has now become possible to cryopreserve hPSCs using conventional cryopreservation methods without any specific equipment. This review summarizes the advances in this area and discusses the optimization of slow cooling cryopreservation for hPSC storage.

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Abbreviations

hPSC:

Human pluripotent stem cell

hESC:

Human embryonic stem cell

hiPSC:

Human induced pluripotent stem cell

DMSO:

Dimethyl sulfoxide

ROCK:

Rho-associated coiled-coil forming kinase

MEF:

Mouse embryonic fibroblast

EG:

Ethylene glycol

KSR:

Knockout serum replacement

FCS:

Fetal calf serum

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Author information

Authors and Affiliations

  1. Institute for Integrated Cell-Material Sciences, Kyoto University, Kyoto, Japan

    Takamichi Miyazaki

  2. Institute for Frontier Medical Sciences, Kyoto University, Kyoto, Japan

    Hirofumi Suemori

Authors
  1. Takamichi Miyazaki
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  2. Hirofumi Suemori
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Correspondence to Hirofumi Suemori .

Editor information

Editors and Affiliations

  1. Department of Oncology, University of Alberta, Edmonton, Alberta, Canada

    Feridoun Karimi-Busheri

  2. Department of Oncology, Cross Cancer Institute and University of Alberta, Edmonton, Alberta, Canada

    Michael Weinfeld

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Miyazaki, T., Suemori, H. (2016). Slow Cooling Cryopreservation Optimized to Human Pluripotent Stem Cells. In: Karimi-Busheri, F., Weinfeld, M. (eds) Biobanking and Cryopreservation of Stem Cells. Advances in Experimental Medicine and Biology, vol 951. Springer, Cham. https://doi.org/10.1007/978-3-319-45457-3_5

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