An antibody against SSEA-5 glycan on human pluripotent stem cells enables removal of teratoma-forming cells

Chad Tang; Andrew S. Lee; Jens Peter Volkmer; Debashis Sahoo; Divya Nag; Adriane R. Mosley; Matthew A. Inlay; Reza Ardehali; Shawn L. Chavez; Renee Reijo Pera; Barry Behr; Joseph C. Wu; Irving L. Weissman; Micha Drukker

doi:10.1038/nbt.1947

An antibody against SSEA-5 glycan on human pluripotent stem cells enables removal of teratoma-forming cells

Chad Tang, Andrew S. Lee, Jens Peter Volkmer, Debashis Sahoo, Divya Nag, Adriane R. Mosley, Matthew A. Inlay, Reza Ardehali, Shawn L. Chavez, Renee Reijo Pera, Barry Behr, Joseph C. Wu, Irving L. Weissman, Micha Drukker

Research output: Contribution to journal › Article › peer-review

313 Scopus citations

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.

Original language	English (US)
Pages (from-to)	829-834
Number of pages	6
Journal	Nature biotechnology
Volume	29
Issue number	9
DOIs	https://doi.org/10.1038/nbt.1947
State	Published - Sep 2011
Externally published	Yes

ASJC Scopus subject areas

Biotechnology
Bioengineering
Applied Microbiology and Biotechnology
Molecular Medicine
Biomedical Engineering

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@article{aadf8819d3e84c92b81e18a92be0912f,

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.",

author = "Chad Tang and Lee, {Andrew S.} and Volkmer, {Jens Peter} and Debashis Sahoo and Divya Nag and Mosley, {Adriane R.} and Inlay, {Matthew A.} and Reza Ardehali and Chavez, {Shawn L.} and Pera, {Renee Reijo} and Barry Behr and Wu, {Joseph C.} and Weissman, {Irving L.} and Micha Drukker",

note = "Funding Information: The authors acknowledge C. Contag for providing luciferase constructs, M. van de Rijn and K. Montgomery for their assistance scanning fetal array slides and providing online access to these slides, P. Chu for assistance with hematoxylin and eosin staining, C. Muscat and T. Naik for assistance with hybridoma culture, W. Zhang for assistance in cell culturing, the Consortium for Functional Glycomics for providing and testing glycan arrays, and T. Serwold and C. Bertozzi for critical advice. This work was supported by funds provided by the California Institute of Regenerative Medicine (CIRM) (Comprehensive grant RC1-00354-1). C.T. and A.S.L. are supported by the Howard Hughes Medical Institute Medical Fellows and the Stanford Medical Scholars Program, J.-P.V. is supported by the Deutsche Forschungsgemeinschaft, C.T., M.A.I., R.A. and M.D. are supported by CIRM (Comprehensive grant RC1-00354-1).",

year = "2011",

month = sep,

doi = "10.1038/nbt.1947",

language = "English (US)",

volume = "29",

pages = "829--834",

journal = "Nature biotechnology",

issn = "1087-0156",

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T1 - An antibody against SSEA-5 glycan on human pluripotent stem cells enables removal of teratoma-forming cells

AU - Tang, Chad

AU - Lee, Andrew S.

AU - Volkmer, Jens Peter

AU - Sahoo, Debashis

AU - Nag, Divya

AU - Mosley, Adriane R.

AU - Inlay, Matthew A.

AU - Ardehali, Reza

AU - Chavez, Shawn L.

AU - Pera, Renee Reijo

AU - Behr, Barry

AU - Wu, Joseph C.

AU - Weissman, Irving L.

AU - Drukker, Micha

N1 - Funding Information: The authors acknowledge C. Contag for providing luciferase constructs, M. van de Rijn and K. Montgomery for their assistance scanning fetal array slides and providing online access to these slides, P. Chu for assistance with hematoxylin and eosin staining, C. Muscat and T. Naik for assistance with hybridoma culture, W. Zhang for assistance in cell culturing, the Consortium for Functional Glycomics for providing and testing glycan arrays, and T. Serwold and C. Bertozzi for critical advice. This work was supported by funds provided by the California Institute of Regenerative Medicine (CIRM) (Comprehensive grant RC1-00354-1). C.T. and A.S.L. are supported by the Howard Hughes Medical Institute Medical Fellows and the Stanford Medical Scholars Program, J.-P.V. is supported by the Deutsche Forschungsgemeinschaft, C.T., M.A.I., R.A. and M.D. are supported by CIRM (Comprehensive grant RC1-00354-1).

PY - 2011/9

Y1 - 2011/9

N2 - 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.

AB - 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.

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JO - Nature biotechnology

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An antibody against SSEA-5 glycan on human pluripotent stem cells enables removal of teratoma-forming cells

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