title: Proteomics-based dissection of human endoderm progenitors by differential cell capture on antibody array.
abstract: Heterogeneity, shortage of material, and lack of progenitor-specific cell surface markers are major obstacles to elucidating the mechanisms underlying developmental processes. Here we report a proteomics platform that alleviates these difficulties and demonstrate its effectiveness in fractionating heterogeneous cultures of early endoderm derived from human embryonic stem cells. The approach, designated differential cell-capture antibody array, is based on highly parallel, comparative screening of live cell populations using hundreds of antibodies directed against cell-surface antigens. We used this platform to fractionate the hitherto unresolved early endoderm compartment of CXCR4+ cells and identify several endoderm (CD61+ and CD63+) and non-endoderm (CD271+, CD49F+, CD44+ and B2M+) sub-populations. We provide evidence that one of these sub-populations, CD61+, is directly derived from CXCR4+ cells, displays characteristic kinetics of emergence, and exhibits a distinct gene expression profile. The results demonstrate the potential of the cell-capture antibody array as a powerful proteomics tool for detailed dissection of heterogeneous cellular systems.
authors.affiliation: Department of Biological Chemistry, Weizmann Institute of Science, Rehovot, 76100, Israel.
authors.forename: Revital
authors.initials: R
authors.lastname: Sharivkin
authors.forename: Michael D
authors.initials: MD
authors.lastname: Walker
authors.forename: Yoav
authors.initials: Y
authors.lastname: Soen
mesh_terms: D000906:Antibodies / Q000276:immunology; D000954:Antigens, Surface / Q000276:immunology*; D015415:Biomarkers / Q000032:analysis; D002454:Cell Differentiation; D002460:Cell Line; D019070:Cell Lineage; D002469:Cell Separation; D053595:Embryonic Stem Cells / Q000737:chemistry* / Q000166:cytology*; D004707:Endoderm / Q000737:chemistry* / Q000166:cytology*; D005434:Flow Cytometry; D006801:Humans; D018960:Hyaluronan Receptors / Q000032:analysis / Q000276:immunology; D039503:Integrin alpha6 / Q000032:analysis / Q000276:immunology; D039661:Integrin beta3 / Q000032:analysis / Q000276:immunology; D009419:Nerve Tissue Proteins / Q000032:analysis / Q000276:immunology; D020411:Oligonucleotide Array Sequence Analysis; D040901:Proteomics; D019718:Receptors, CXCR4 / Q000032:analysis* / Q000276:immunology; D017475:Receptors, Nerve Growth Factor / Q000032:analysis / Q000276:immunology; D060149:Tetraspanin 30 / Q000032:analysis
chemical_list: D000906:Antibodies; D000954:Antigens, Surface; D015415:Biomarkers; C497483:CD44 protein, human; C554586:CD63 protein, human; C527276:CXCR4 protein, human; D018960:Hyaluronan Receptors; D039503:Integrin alpha6; D039661:Integrin beta3; C484065:NGFR protein, human; D009419:Nerve Tissue Proteins; D019718:Receptors, CXCR4; D017475:Receptors, Nerve Growth Factor; D060149:Tetraspanin 30