title: Resolving Heterogeneity: Fluorescence-Activated Cell Sorting of Dynamic Cell Populations from Feeder-Free Mouse Embryonic Stem Cell Culture.
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).
authors.affiliation: The Danish Stem Cell Centre-DanStem, University of Copenhagen, 3B Blegdamsvej, DK-2200, Copenhagen, Denmark.
authors.forename: Jurriaan
authors.initials: J
authors.lastname: Hölzenspies
authors.affiliation: The Danish Stem Cell Centre-DanStem, University of Copenhagen, 3B Blegdamsvej, DK-2200, Copenhagen, Denmark.
authors.forename: Gelo
authors.initials: G
authors.lastname: Dela Cruz
authors.affiliation: The Danish Stem Cell Centre-DanStem, University of Copenhagen, 3B Blegdamsvej, DK-2200, Copenhagen, Denmark. Joshua.brickman@sund.ku.dk.
authors.forename: Joshua
authors.initials: J
authors.lastname: M Brickman
mesh_terms: D000818:Animals; D018929:Cell Culture Techniques / Q000379:methods; D002454:Cell Differentiation; D019070:Cell Lineage; D049109:Cell Proliferation; D004707:Endoderm / Q000166:cytology; D005434:Flow Cytometry / Q000379:methods*; D051379:Mice; D000066450:Mouse Embryonic Stem Cells / Q000166:cytology* / Q000378:metabolism; D011506:Proteins / Q000235:genetics / Q000302:isolation & purification; D012313:RNA / Q000235:genetics / Q000302:isolation & purification; D014158:Transcription, Genetic
chemical_list: D011506:Proteins; D012313:RNA
keywords: Embryonic stem cells; Endoderm; Lineage priming; Pluripotency; Self renewal; Transcription