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Staining Embryonic Stem Cells Using Monoclonal Antibodies to Stage-Specific Embryonic Antigens

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Nuclear Reprogramming

Part of the book series: Methods in Molecular Biology™ ((MIMB,volume 325))

Abstract

Stage-specific embryonic antigens (SSEAs) are cell-surface molecules that exhibit lineage-restricted patterns of expression during development. These antigens provide useful markers for identifying embryonic stem cells and their differentiated derivatives. SSEA-1 provides a surface marker for mouse and human primordial germ cells and mouse embryonic stem cells. SSEA-3 and SSEA-4 provide surface markers for human embryonic stem cells. Here, we describe methods for staining embryonic stem cells in culture and suspension using specific primary antibodies and fluorescent dye- or enzyme-conjugated secondary antibodies. Methods for alkaline phosphatase staining and immunohistochemical localization of antigens in paraffin sections are also described. Monoclonal antibodies directed to SSEAs are valuable research tools that can be used to: (1) determine the percentage of stem cells in mixed cultures; (2) examine the spatial distribution of stem cells and differentiated derivatives in monolayer cultures and embryoid bodies; and (3) isolate populations of differentiated and undifferentiated cells by immunomagnetic or fluorescence-activated cell sorting for cellular and molecular analyses.

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Author information

Authors and Affiliations

  1. Department of Pathology, Anatomy, and Cell Biology, Thomas Jefferson University, USA

    Bruce A. Fenderson

  2. Jefferson Medical College, Philadelphia, PA

    Bruce A. Fenderson

  3. Cell Therapy Laboratory, La Paz Hospital, Madrid, Spain

    Maria P. De Miguel

  4. Institute for Cell Engineering, The Johns Hopkins University, Baltimore, MD

    Maria P. De Miguel & Peter J. Donovan

  5. Institute for Cell Engineering, The Johns Hopkins University, Baltimore, MD

    April D. Pyle

Authors
  1. Bruce A. Fenderson
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  2. Maria P. De Miguel
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  3. April D. Pyle
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  4. Peter J. Donovan
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Editor information

Editors and Affiliations

  1. Department of Gene Function and Development, Roslin Institute, Roslin, Midlothian, UK

    Steve Pells

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© 2006 Humana Press Inc.

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Fenderson, B.A., De Miguel, M.P., Pyle, A.D., Donovan, P.J. (2006). Staining Embryonic Stem Cells Using Monoclonal Antibodies to Stage-Specific Embryonic Antigens. In: Pells, S. (eds) Nuclear Reprogramming. Methods in Molecular Biology™, vol 325. Humana Press. https://doi.org/10.1385/1-59745-005-7:207

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  • DOI: https://doi.org/10.1385/1-59745-005-7:207

  • Publisher Name: Humana Press

  • Print ISBN: 978-1-58829-379-4

  • Online ISBN: 978-1-59745-005-8

  • eBook Packages: Springer Protocols

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