| RRC ID |
37493
|
| Author |
Imaizumi K, Iha M, Nishishita N, Kawamata S, Nishikawa S, Akuta T.
|
| Title |
A Simple and Efficient Method of Slow Freezing for Human Embryonic Stem Cells and Induced Pluripotent Stem Cells.
|
| Journal |
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.
|
| Volume |
1341
|
| Pages |
15-24
|
| Published |
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
|
| Resource |
| Human and Animal Cells |
201B7(HPS0063)
KhES-1(HES0001) |
