Embryonic Stem Cells/Induced Pluripotent Stem Cells

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Characterization of Epithelial Cell Adhesion Molecule as a Surface Marker on Undifferentiated Human Embryonic Stem Cells†‡§

  1. Valerie Y. Ng1,¶,*,
  2. Sheu Ngo Ang1,
  3. Jia Xin Chan1,
  4. Andre B.H. Choo1,2

Article first published online: 25 SEP 2009

DOI: 10.1002/stem.221

Issue

STEM CELLS

STEM CELLS

Volume 28, Issue 1, pages 29–35, January 2010

Additional Information

How to Cite

Ng, V. Y., Ang, S. N., Chan, J. X. and Choo, A. B. (2010), Characterization of Epithelial Cell Adhesion Molecule as a Surface Marker on Undifferentiated Human Embryonic Stem Cells. STEM CELLS, 28: 29–35. doi: 10.1002/stem.221

Author Information

  1. 1

    Bioprocessing Technology Institute, Agency for Science, Technology and Research (A*STAR), Biopolis, Singapore

  2. 2

    Division of Bioengineering, Faculty of Engineering, National University of Singapore, Singapore

  1. Telephone: 65-6407-0843; Fax: 65-6478-9561

*Stem Cell Group, Bioprocessing Technology Institute, 20 Biopolis Way, 06-01, Centros, Singapore 138668, Singapore

  1. †

    Author contributions: V.Y.N.: conception and design, collection and/or assembly of data, data analysis and interpretation, manuscript writing; S.N.A. and J.X.C.: collection and/or assembly of data; A.B.H.C.: conception and design, final approval of manuscript.

  2. ‡

    Disclosure of potential conflicts of interest is found at the end of this article.

  3. §

    First published online in STEM CELLS EXPRESS September 25, 2009.

Publication History

  1. Issue published online: 12 JAN 2010
  2. Article first published online: 25 SEP 2009
  3. Accepted manuscript online: 25 SEP 2009 12:00AM EST
  4. Manuscript Accepted: 3 SEP 2009
  5. Manuscript Received: 28 APR 2009

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Keywords:

  • Embryonic stem cells;
  • Monoclonal antibodies;
  • Cell surface markers;
  • Cell adhesion molecules

Abstract

Human embryonic stem cells (hESCs) have the capacity to remain pluripotent and self-renew indefinitely. To discover novel players in the maintenance of hESCs, we have previously reported the generation of monoclonal antibodies that bind to cell surface markers on hESCs, and not to mouse embryonic stem cells or differentiated embryoid bodies. In this study, we have identified the antigen target of one such monoclonal antibody as the epithelial cell adhesion molecule (EpCAM). In undifferentiated hESCs, EpCAM is localized to Octamer 4 (OCT4)-positive pluripotent cells, and its expression is down-regulated upon differentiation. To further understand its biological function in hESCs, endogenous EpCAM expression was silenced using small interfering RNA. EpCAM knockdown had marginal negative effects on OCT4 and TRA-1-60 expression, however cell proliferation was decreased by >40%. Examination of lineage marker expression showed marked upregulation of endoderm and mesoderm genes in EpCAM-silenced cells, under both pluripotent and differentiating conditions. These results were validated using a hESC line whose EpCAM expression has been stably knocked down. Data from the stable line confirmed that downregulation of EpCAM decreases cell growth and increases gene expression in the endoderm and mesoderm lineages. In vivo, hESCs lacking EpCAM were able to form teratomas containing tissues representing the three germ layers, and gene expression analysis yielded marked increase in the endoderm marker alpha fetoprotein compared with control. Together these data demonstrate that EpCAM is a surface marker on undifferentiated hESCs and plays functional roles in proliferation and differentiation. STEM CELLS 2010;28:29–35

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