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Published in:

2015 | Online First | Chapter

A Simple and Efficient Method of Slow Freezing for Human Embryonic Stem Cells and Induced Pluripotent Stem Cells

Authors : Keitaro Imaizumi, Momoe Iha, Naoki Nishishita, Shin Kawamata, M.D., Ph.D., Shinichi Nishikawa, Teruo Akuta, Ph.D.

Published in: Methods in Molecular Biology™

Publisher: Springer New York

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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.
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Metadata
Title
A Simple and Efficient Method of Slow Freezing for Human Embryonic Stem Cells and Induced Pluripotent Stem Cells
Authors
Keitaro Imaizumi
Momoe Iha
Naoki Nishishita
Shin Kawamata, M.D., Ph.D.
Shinichi Nishikawa
Teruo Akuta, Ph.D.
Copyright Year
2015
Publisher
Springer New York
DOI
https://doi.org/10.1007/7651_2015_211