| GeneLibs

题目:: StemCell Keep™ is effective for cryopreservation of human embryonic stem cells by vitrification.
作者:: Ota(Akemi),Matsumura(Kazuaki),Lee(Jun-Jae),Sumi(Shoichiro),Hyon(Soung Hyu)
状态:: 发布时间2016-08-09 , 更新时间 2016-08-09
期刊:: Cell Transplant
摘要:: Safe and stable cryopreservation is critical for research involving human embryonic stem (hES) cells. 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 (hiPS) cells by vitrification. Because hES and hiPS cells are not identical, the current study examined the use of SCK on hES cells. hES cells 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 hES cells were assessed for their attachment ability, and characterized by alkalinephosphatase (AP) and immunocytochemical (ICC) staining, fluorescence activated cell sorting (FACS), the reverse transcription polymerase chain reaction (RT-PCR), and karyotyping. The proliferation of SCK-cryopreserved hES cells cultured on SNL cells, or in feeder-free conditions, was higher than that of cells preserved with DAP213 (2 M DMSO, 1 M acetamide, and 3 M propylene glycol). The cell number with SCK-cryopreserved hES cells was about twice that of hES cells cryopreserved in DAP213. The pluripotency of SCK-cryopreserved hES cells was similar to that of DAP213-cryopreserved hES cells based on AP staining. Data from ICC, FACS, and RTPCR analyses showed that stem cell markers were continually expressed on SCK-cryopreserved hES cells. The teratoma assay showed that SCK-cryopreserved hES cells differentiated into three germ layers. Furthermore, SCK-cryopreserved hES cells had normal karyotypes. These data indicated that SCK was effective for cryopreservation of hES cells by vitrification.
语言:: eng
DOI:: 10.3727/096368916X692654

文献库文献相关信息

DataTable pData

联系方式

微信公众号

文献库 文献相关信息

DataTable pData

联系方式

微信公众号

文献库文献相关信息