A cell surfaceome map for immunophenotyping and sorting pluripotent stem cells

Rebekah L. Gundry, Daniel R. Riordon, Yelena Tarasova, Sandra Chuppa, Subarna Bhattacharya, Ondrej Juhasz, Olena Wiedemeier, Samuel Milanovich, Fallon K. Noto, Irina Tchernyshyov, Kimberly Raginski, Damaris Bausch-Fluck, Hyun Jin Tae, Shannon Marshall, Stephen A. Duncan, Bernd Wollscheid, Robert P. Wersto, Sridhar Rao, Jennifer E. Van Eyk, Kenneth R. Boheler

Research output: Contribution to journalArticlepeer-review

37 Scopus 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.

Original languageEnglish (US)
Pages (from-to)303-316
Number of pages14
JournalMolecular and Cellular Proteomics
Volume11
Issue number8
DOIs
StatePublished - Aug 2012

ASJC Scopus subject areas

  • Analytical Chemistry
  • Biochemistry
  • Molecular Biology

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