title: Cell-surface proteomics identifies lineage-specific markers of embryo-derived stem cells.
abstract: The advent of reprogramming and its impact on stem cell biology has renewed interest in lineage restriction in mammalian embryos, the source of embryonic (ES), epiblast (EpiSC), trophoblast (TS), and extraembryonic endoderm (XEN) stem cell lineages. Isolation of specific cell types during stem cell differentiation and reprogramming, and also directly from embryos, is a major technical challenge because few cell-surface proteins are known that can distinguish each cell type. We provide a large-scale proteomic resource of cell-surface proteins for the four embryo-derived stem cell lines. We validated 27 antibodies against lineage-specific cell-surface markers, which enabled investigation of specific cell populations during ES-EpiSC reprogramming and ES-to-XEN differentiation. Identified markers also allowed prospective isolation and characterization of viable lineage progenitors from blastocysts by flow cytometry. These results provide a comprehensive stem cell proteomic resource and enable new approaches to interrogate the mechanisms that regulate cell fate specification.
authors.affiliation: Program in Developmental and Stem Cell Biology, Hospital for Sick Children Research Institute, 555 University Avenue, Toronto, ON M5G 1X8, Canada.
authors.forename: Peter J
authors.initials: PJ
authors.lastname: Rugg-Gunn
authors.forename: Brian J
authors.initials: BJ
authors.lastname: Cox
authors.forename: Fredrik
authors.initials: F
authors.lastname: Lanner
authors.forename: Parveen
authors.initials: P
authors.lastname: Sharma
authors.forename: Vladimir
authors.initials: V
authors.lastname: Ignatchenko
authors.forename: Angela C H
authors.initials: AC
authors.lastname: McDonald
authors.forename: Jodi
authors.initials: J
authors.lastname: Garner
authors.forename: Anthony O
authors.initials: AO
authors.lastname: Gramolini
authors.forename: Janet
authors.initials: J
authors.lastname: Rossant
authors.forename: Thomas
authors.initials: T
authors.lastname: Kislinger
mesh_terms: D000818:Animals; D015415:Biomarkers / Q000378:metabolism*; D001755:Blastocyst / Q000166:cytology / Q000378:metabolism; D015153:Blotting, Western; D002454:Cell Differentiation; D019070:Cell Lineage; D002478:Cells, Cultured; D004622:Embryo, Mammalian / Q000166:cytology / Q000378:metabolism*; D053595:Embryonic Stem Cells / Q000166:cytology / Q000378:metabolism*; D004707:Endoderm / Q000166:cytology / Q000378:metabolism*; D005434:Flow Cytometry; D020869:Gene Expression Profiling; D018507:Gene Expression Regulation, Developmental; D051379:Mice; D020411:Oligonucleotide Array Sequence Analysis; D040901:Proteomics; D012333:RNA, Messenger / Q000235:genetics; D060888:Real-Time Polymerase Chain Reaction; D014327:Trophoblasts / Q000166:cytology / Q000378:metabolism*
chemical_list: D015415:Biomarkers; D012333:RNA, Messenger