Resolving Heterogeneity: Fluorescence-Activated Cell Sorting of Dynamic Cell Populations from Feeder-Free Mouse Embryonic Stem Cell Culture

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Abstract

Embryonic stem cell (ESC) culture comprises a mixture of cells that are primed to differentiate into different lineages. In conditions where ESCs self-renew, these primed populations continuously interconvert and consequently show highly dynamic coordinated changes in their expression of different sets of pluripotency and differentiation markers. It has become increasingly apparent that this transcriptional heterogeneity is an important characteristic of ESC culture. By sorting for specific populations of ESCs it is possible to enrich for cells with a capacity to colonize the embryo proper or the extra-embryonic lineages such as the descendents of the primitive endoderm or trophoblast. Here, we describe a method of isolating specific sub-sets of ESCs from the pluripotent cells present in in vitro ESC culture using SSEA1 antibody staining in combination with reporter lines and fluorescence activated cell sorting (FACS).

OriginalsprogEngelsk
TitelEmbryonic Stem Cell Protocols
RedaktørerKursad Turksen
Antal sider16
Vol/bind1341
ForlagSpringer
Publikationsdato1 okt. 2016
Sider25-40
ISBN (Trykt)978-1-4939-2953-5
ISBN (Elektronisk)978-1-4939-2953-5
DOI
StatusUdgivet - 1 okt. 2016
NavnMethods in Molecular Biology
Vol/bind134
ISSN1064-3745

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