title: An antibody against SSEA-5 glycan on human pluripotent stem cells enables removal of teratoma-forming cells.
abstract: An important risk in the clinical application of human pluripotent stem cells (hPSCs), including human embryonic and induced pluripotent stem cells (hESCs and hiPSCs), is teratoma formation by residual undifferentiated cells. We raised a monoclonal antibody against hESCs, designated anti-stage-specific embryonic antigen (SSEA)-5, which binds a previously unidentified antigen highly and specifically expressed on hPSCs--the H type-1 glycan. Separation based on SSEA-5 expression through fluorescence-activated cell sorting (FACS) greatly reduced teratoma-formation potential of heterogeneously differentiated cultures. To ensure complete removal of teratoma-forming cells, we identified additional pluripotency surface markers (PSMs) exhibiting a large dynamic expression range during differentiation: CD9, CD30, CD50, CD90 and CD200. Immunohistochemistry studies of human fetal tissues and bioinformatics analysis of a microarray database revealed that concurrent expression of these markers is both common and specific to hPSCs. Immunodepletion with antibodies against SSEA-5 and two additional PSMs completely removed teratoma-formation potential from incompletely differentiated hESC cultures.
authors.affiliation: Institute of Stem Cell Biology and Regenerative Medicine, Stanford University School of Medicine, Stanford, California, USA.
authors.forename: Chad
authors.initials: C
authors.lastname: Tang
authors.forename: Andrew S
authors.initials: AS
authors.lastname: Lee
authors.forename: Jens-Peter
authors.initials: JP
authors.lastname: Volkmer
authors.forename: Debashis
authors.initials: D
authors.lastname: Sahoo
authors.forename: Divya
authors.initials: D
authors.lastname: Nag
authors.forename: Adriane R
authors.initials: AR
authors.lastname: Mosley
authors.forename: Matthew A
authors.initials: MA
authors.lastname: Inlay
authors.forename: Reza
authors.initials: R
authors.lastname: Ardehali
authors.forename: Shawn L
authors.initials: SL
authors.lastname: Chavez
authors.forename: Renee Reijo
authors.initials: RR
authors.lastname: Pera
authors.forename: Barry
authors.initials: B
authors.lastname: Behr
authors.forename: Joseph C
authors.initials: JC
authors.lastname: Wu
authors.forename: Irving L
authors.initials: IL
authors.lastname: Weissman
authors.forename: Micha
authors.initials: M
authors.lastname: Drukker
mesh_terms: D000818:Animals; D000911:Antibodies, Monoclonal / Q000378:metabolism*; D015415:Biomarkers; D002454:Cell Differentiation; D002478:Cells, Cultured; D019295:Computational Biology; D005434:Flow Cytometry; D005786:Gene Expression Regulation; D006801:Humans; D007150:Immunohistochemistry; D051379:Mice; D008807:Mice, Inbred BALB C; D046228:Microarray Analysis; D039904:Pluripotent Stem Cells / Q000737:chemistry / Q000166:cytology*; D011134:Polysaccharides / Q000378:metabolism*; D060888:Real-Time Polymerase Chain Reaction; D055746:Stage-Specific Embryonic Antigens / Q000378:metabolism*; D013724:Teratoma / Q000378:metabolism / Q000473:pathology*
chemical_list: D000911:Antibodies, Monoclonal; D015415:Biomarkers; D011134:Polysaccharides; D055746:Stage-Specific Embryonic Antigens