RRC ID 37493
著者 Imaizumi K, Iha M, Nishishita N, Kawamata S, Nishikawa S, Akuta T.
タイトル A Simple and Efficient Method of Slow Freezing for Human Embryonic Stem Cells and Induced Pluripotent Stem Cells.
ジャーナル Methods Mol Biol
Abstract Protocols available for the cryopreservation of human embryonic stem (ES) and induced pluripotent stem (iPS) cells are very inefficient and laborious compared to those for the cryopreservation of murine ES/iPS cells or other general cell lines. While the vitrification method may be adequate when working with small numbers of human ES/iPS cells, it requires special skills and is unsuitable when working with large cell numbers. Here, we describe a simple and efficient method for the cryopreservation of hES/hiPS cells that is based on a conventional slow freezing method that uses a combination of Pronase/EDTA for Stem™ and CP-5E™ [final concentrations: 6 % hydroxyethyl starch, 5 % DMSO, and 5 % ethylene glycol in saline]. CP-5E™ is highly effective for the cryopreservation of small cell clumps produced by hES/hiPS colony detachment in the presence of Pronase and EDTA (Pronase/EDTA for Stem™, a formulation containing multiple digestive enzymes from Streptomyces griseus). This novel method would be quite useful for large-scale hES/iPS cell banking for use in clinical applications.
巻・号 1341
ページ 15-24
公開日 2016-1-1
DOI 10.1007/7651_2015_211
PMID 26069023
MeSH Cell Culture Techniques / methods Cell Proliferation Cryopreservation / economics Cryopreservation / methods* Cryoprotective Agents / pharmacology* Dimethyl Sulfoxide / pharmacology Ethylene Glycol / pharmacology Freezing Human Embryonic Stem Cells / cytology* Human Embryonic Stem Cells / drug effects Humans Hydroxyethyl Starch Derivatives / pharmacology Induced Pluripotent Stem Cells / cytology* Induced Pluripotent Stem Cells / drug effects Pronase / pharmacology Staining and Labeling / methods Streptomyces griseus / enzymology
リソース情報
ヒト・動物細胞 201B7(HPS0063) KhES-1(HES0001)