Chapter title |
A Simple and Efficient Method of Slow Freezing for Human Embryonic Stem Cells and Induced Pluripotent Stem Cells.
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Chapter number | 211 |
Book title |
Embryonic Stem Cell Protocols
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Published in |
Methods in molecular biology, June 2015
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DOI | 10.1007/7651_2015_211 |
Pubmed ID | |
Book ISBNs |
978-1-4939-2953-5, 978-1-4939-2954-2
|
Authors |
Imaizumi, Keitaro, Iha, Momoe, Nishishita, Naoki, Kawamata, Shin, Nishikawa, Shinichi, Akuta, Teruo, Keitaro Imaizumi, Momoe Iha, Naoki Nishishita, Shin Kawamata, Shinichi Nishikawa, Teruo Akuta |
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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