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A cell surfaceome map for immunophenotyping and sorting pluripotent stem cells. Mol Cell Proteomics 2012 Aug;11(8):303-16

Date

04/12/2012

Pubmed ID

22493178

Pubmed Central ID

PMC3412963

DOI

10.1074/mcp.M112.018135

Scopus ID

2-s2.0-84864821878 (requires institutional sign-in at Scopus site)   52 Citations

Abstract

Induction of a pluripotent state in somatic cells through nuclear reprogramming has ushered in a new era of regenerative medicine. Heterogeneity and varied differentiation potentials among induced pluripotent stem cell (iPSC) lines are, however, complicating factors that limit their usefulness for disease modeling, drug discovery, and patient therapies. Thus, there is an urgent need to develop nonmutagenic rapid throughput methods capable of distinguishing among putative iPSC lines of variable quality. To address this issue, we have applied a highly specific chemoproteomic targeting strategy for de novo discovery of cell surface N-glycoproteins to increase the knowledge-base of surface exposed proteins and accessible epitopes of pluripotent stem cells. We report the identification of 500 cell surface proteins on four embryonic stem cell and iPSCs lines and demonstrate the biological significance of this resource on mouse fibroblasts containing an oct4-GFP expression cassette that is active in reprogrammed cells. These results together with immunophenotyping, cell sorting, and functional analyses demonstrate that these newly identified surface marker panels are useful for isolating iPSCs from heterogeneous reprogrammed cultures and for isolating functionally distinct stem cell subpopulations.

Author List

Gundry RL, Riordon DR, Tarasova Y, Chuppa S, Bhattacharya S, Juhasz O, Wiedemeier O, Milanovich S, Noto FK, Tchernyshyov I, Raginski K, Bausch-Fluck D, Tae HJ, Marshall S, Duncan SA, Wollscheid B, Wersto RP, Rao S, Van Eyk JE, Boheler KR

Author

Sridhar Rao MD, PhD Associate Professor in the Pediatrics department at Medical College of Wisconsin




MESH terms used to index this publication - Major topics in bold

Animals
Cell Separation
Cells, Cultured
Cytokine Receptor gp130
Embryo, Mammalian
Embryonic Stem Cells
Fibroblasts
Flow Cytometry
Glycoproteins
Green Fluorescent Proteins
Immunophenotyping
Induced Pluripotent Stem Cells
Mass Spectrometry
Membrane Proteins
Mice
Mice, 129 Strain
Mice, Transgenic
Microscopy, Confocal
Octamer Transcription Factor-3
Pluripotent Stem Cells
Proteomics
Teratoma