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Embryonic and induced pluripotent stem cell staining and sorting with the live-cell fluorescence imaging probe CDy1

Authors :
Hyung-Ho Ha
Sungjin Park
Nam-Young Kang
Seong-Wook Yun
Young-Tae Chang
Source :
Nature Protocols. 6:1044-1052
Publication Year :
2011
Publisher :
Springer Science and Business Media LLC, 2011.

Abstract

Detecting and isolating specific types of cells is crucial to understanding a variety of biological processes, including development, aging, regeneration and pathogenesis; this understanding, in turn, allows the use of cells for therapeutic purposes, for which stem cells have emerged recently as invaluable materials. The current methods of isolation and characterization of stem cells depend on cell morphology in culture or on immunostaining of specific markers. These methods are, however, time consuming and involve the use of antibodies that may often make the cells unsuitable for further study. We recently developed a fluorescent small molecule named CDy1 (compound of designation yellow 1) that selectively stains live embryonic stem cells (ESCs) and induced pluripotent stem cells (iPSCs). This protocol describes detailed procedures for staining ESC and iPSC in live conditions and for fluorescence-activated cell sorting (FACS) of ESC using CDy1. Cell staining, image acquisition and FACS can be done within 6 h.

Details

ISSN :
17502799 and 17542189
Volume :
6
Database :
OpenAIRE
Journal :
Nature Protocols
Accession number :
edsair.doi.dedup.....7c1a5601db34503ba55624ed505f2229