title: A cell surfaceome map for immunophenotyping and sorting pluripotent stem cells.
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.
authors.affiliation: Department of Biochemistry, Medical College of Wisconsin, Milwaukee, Wisconsin 53226, USA.
authors.forename: Rebekah L
authors.initials: RL
authors.lastname: Gundry
authors.forename: Daniel R
authors.initials: DR
authors.lastname: Riordon
authors.forename: Yelena
authors.initials: Y
authors.lastname: Tarasova
authors.forename: Sandra
authors.initials: S
authors.lastname: Chuppa
authors.forename: Subarna
authors.initials: S
authors.lastname: Bhattacharya
authors.forename: Ondrej
authors.initials: O
authors.lastname: Juhasz
authors.forename: Olena
authors.initials: O
authors.lastname: Wiedemeier
authors.forename: Samuel
authors.initials: S
authors.lastname: Milanovich
authors.forename: Fallon K
authors.initials: FK
authors.lastname: Noto
authors.forename: Irina
authors.initials: I
authors.lastname: Tchernyshyov
authors.forename: Kimberly
authors.initials: K
authors.lastname: Raginski
authors.forename: Damaris
authors.initials: D
authors.lastname: Bausch-Fluck
authors.forename: Hyun-Jin
authors.initials: HJ
authors.lastname: Tae
authors.forename: Shannon
authors.initials: S
authors.lastname: Marshall
authors.forename: Stephen A
authors.initials: SA
authors.lastname: Duncan
authors.forename: Bernd
authors.initials: B
authors.lastname: Wollscheid
authors.forename: Robert P
authors.initials: RP
authors.lastname: Wersto
authors.forename: Sridhar
authors.initials: S
authors.lastname: Rao
authors.forename: Jennifer E
authors.initials: JE
authors.lastname: Van Eyk
authors.forename: Kenneth R
authors.initials: KR
authors.lastname: Boheler
mesh_terms: D000818:Animals; D002469:Cell Separation / Q000379:methods*; D002478:Cells, Cultured; D050822:Cytokine Receptor gp130 / Q000032:analysis; D004622:Embryo, Mammalian / Q000166:cytology; D053595:Embryonic Stem Cells / Q000166:cytology / Q000378:metabolism / Q000637:transplantation; D005347:Fibroblasts / Q000166:cytology / Q000378:metabolism; D005434:Flow Cytometry; D006023:Glycoproteins / Q000032:analysis*; D049452:Green Fluorescent Proteins / Q000235:genetics / Q000378:metabolism; D016130:Immunophenotyping / Q000379:methods*; D057026:Induced Pluripotent Stem Cells / Q000166:cytology / Q000378:metabolism / Q000637:transplantation; D013058:Mass Spectrometry; D008565:Membrane Proteins / Q000032:analysis*; D051379:Mice; D057507:Mice, 129 Strain; D008822:Mice, Transgenic; D018613:Microscopy, Confocal; D050814:Octamer Transcription Factor-3 / Q000235:genetics / Q000378:metabolism; D039904:Pluripotent Stem Cells / Q000166:cytology / Q000378:metabolism*; D040901:Proteomics / Q000379:methods*; D013724:Teratoma / Q000378:metabolism / Q000473:pathology
chemical_list: D006023:Glycoproteins; D008565:Membrane Proteins; D050814:Octamer Transcription Factor-3; C494174:Pou5f1 protein, mouse; D050822:Cytokine Receptor gp130; D049452:Green Fluorescent Proteins