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Please use this identifier to cite or link to this item: http://hdl.handle.net/10119/14208

Title: StemCell Keep^TM Is Effective for Cryopreservation of Human Embryonic Stem Cells by Vitrification
Authors: Ota, Akemi
Matsumura, Kazuaki
Lee, Jun-Jae
Sumi, Shoichiro
Hyon, Soung-Hyu
Keywords: Cryopreservation
Human embryonic stem cells (hESCs)
Vitrification
Dimethyl sulfoxide free
Polyampholytes
Issue Date: 2017-05-09
Publisher: Cognizant Communication Corporation
Magazine name: Cell Transplantation
Volume: 26
Number: 5
Start page: 773
End page: 787
DOI: 10.3727/096368916X692654
Abstract: Safe and stable cryopreservation is critical for research involving human embryonic stem cells (hESCs). Dimethyl sulfoxide (DMSO) is a popular cryoprotective agent; however, its cytotoxicity cannot be ignored. Thus, there is a need for an alternate cryoprotectant. We reported previously that a novel cryopreservation reagent, StemCell Keep™ (SCK), was effective for cryopreserving human induced pluripotent stem cells (hiPSCs) by vitrification. Because hESCs and hiPSCs are not identical, the current study examined the use of SCK on hESCs. hESCs cryopreserved with SCK were thawed and cultured on SNL 76/7 cells, which were derived from a mouse fibroblast STO cell line transformed with neomycin resistance and murine LIF genes. After cryopreservation, cultured hESCs were assessed for their attachment ability and characterized by alkaline phosphatase (AP) and immunocytochemical (ICC) staining, fluorescence-activated cell sorting (FACS), reverse transcription polymerase chain reaction (RT-PCR), and karyotyping. The proliferation of SCK-cryopreserved hESCs cultured on SNL cells, or in feeder-free conditions, was higher than that of cells preserved in a solution of 2 M DMSO, 1 M acetamide, and 3 M propylene glycol (DAP). The cell number with SCK-cryopreserved hESCs was about twice that of hESCs cryopreserved in DAP. The pluripotency of SCK-cryopreserved hESCs was similar to that of DAP-cryopreserved hESCs based on AP staining. Data from ICC, FACS, and RT-PCR analyses showed that stem cell markers were continually expressed on SCK-cryopreserved hESCs. The teratoma assay showed that SCK-cryopreserved hESCs differentiated into three germ layers. Furthermore, SCK-cryopreserved hESCs had normal karyotypes. These data indicate that SCK was effective for cryopreservation of hESCs by vitrification.
Rights: Copyright (C) 2017 Cognizant, LLC. This is an open access article under the terms of the Creative Commons Attribution Non-Commercial (CC BY NC) license. https://creativecommons.org/licenses/by-nc/3.0/
URI: http://hdl.handle.net/10119/14208
Material Type: publisher
Appears in Collections:c10-1. 雑誌掲載論文 (Journal Articles)

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