Generation of monoclonal antibodies specific for cell surface molecules expressed on early mouse endoderm

Paul Gadue, Valerie Gouon-Evans, Xin Cheng, Ewa Wandzioch, Kenneth S. Zaret, Markus Grompe, Philip R. Streeter, Gordon M. Keller

Research output: Contribution to journalArticle

31 Scopus citations

Abstract

The development of functional cell populations such as hepatocytes and pancreatic β cells from embryonic stem cell (ESC) is dependent on the efficient induction of definitive endoderm early in the differentiation process. To monitor definitive endoderm formation in mouse ESC differentiation cultures in a quantitative fashion, we generated a reporter cell line that expresses human CD25 from the Foxa3 locus and human CD4 from the Foxa2 locus. Induction of these reporter ESCs with high concentrations of activin A led to the development of a CD25-Foxa3+CD4-Foxa2+ population within 4-5 days of culture. Isolation and characterization of this population showed that it consists predominantly of definitive endoderm that is able to undergo hepatic specification under the appropriate conditions. To develop reagents that can be used for studies on endoderm development from unmanipulated ESCs, from induced pluripotent stem cells, and from the mouse embryo, we generated monoclonal antibodies against the CD25-Foxa3+CD4-Foxa2+ population. With this approach, we identified two antibodies that react specifically with endoderm from ESC cultures and from the early embryo. The specificity of these antibodies enables one to quantitatively monitor endoderm development in ESC differentiation cultures, to study endoderm formation in the embryo, and to isolate pure populations of culture- or embryo-derived endodermal cells.

Original languageEnglish (US)
Pages (from-to)2103-2113
Number of pages11
JournalStem Cells
Volume27
Issue number9
DOIs
StatePublished - Sep 2009

Keywords

  • Differentiation
  • Differentiation antigens
  • EB
  • ESC
  • Monoclonal antibodies

ASJC Scopus subject areas

  • Molecular Medicine
  • Developmental Biology
  • Cell Biology

Fingerprint Dive into the research topics of 'Generation of monoclonal antibodies specific for cell surface molecules expressed on early mouse endoderm'. Together they form a unique fingerprint.

  • Cite this

    Gadue, P., Gouon-Evans, V., Cheng, X., Wandzioch, E., Zaret, K. S., Grompe, M., Streeter, P. R., & Keller, G. M. (2009). Generation of monoclonal antibodies specific for cell surface molecules expressed on early mouse endoderm. Stem Cells, 27(9), 2103-2113. https://doi.org/10.1002/stem.147