TY - JOUR
T1 - A cell surfaceome map for immunophenotyping and sorting pluripotent stem cells
AU - Gundry, Rebekah L.
AU - Riordon, Daniel R.
AU - Tarasova, Yelena
AU - Chuppa, Sandra
AU - Bhattacharya, Subarna
AU - Juhasz, Ondrej
AU - Wiedemeier, Olena
AU - Milanovich, Samuel
AU - Noto, Fallon K.
AU - Tchernyshyov, Irina
AU - Raginski, Kimberly
AU - Bausch-Fluck, Damaris
AU - Tae, Hyun Jin
AU - Marshall, Shannon
AU - Duncan, Stephen A.
AU - Wollscheid, Bernd
AU - Wersto, Robert P.
AU - Rao, Sridhar
AU - Van Eyk, Jennifer E.
AU - Boheler, Kenneth R.
PY - 2012/8
Y1 - 2012/8
N2 - 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.
AB - 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.
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U2 - 10.1074/mcp.M112.018135
DO - 10.1074/mcp.M112.018135
M3 - Article
C2 - 22493178
AN - SCOPUS:84864821878
SN - 1535-9476
VL - 11
SP - 303
EP - 316
JO - Molecular and Cellular Proteomics
JF - Molecular and Cellular Proteomics
IS - 8
ER -