Abrogation of E-Cadherin-Mediated Cell–Cell Contact in Mouse Embryonic Stem Cells Results in Reversible LIF-Independent Self-Renewal

Authors

  • Francesca Soncin,

    1. Core Technology Facility, Faculty of Medical and Human Sciences, and The University of Manchester, Manchester, United Kingdom
    2. Materials Science Centre, The University of Manchester, Manchester, United Kingdom
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  • Lisa Mohamet,

    1. Core Technology Facility, Faculty of Medical and Human Sciences, and The University of Manchester, Manchester, United Kingdom
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  • Dominik Eckardt,

    1. Department of Molecular Embryology, Max-Planck Institute of Immunobiology, Freiburg, Germany
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  • Sarah Ritson,

    1. Core Technology Facility, Faculty of Medical and Human Sciences, and The University of Manchester, Manchester, United Kingdom
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  • Angela M. Eastham,

    1. Core Technology Facility, Faculty of Medical and Human Sciences, and The University of Manchester, Manchester, United Kingdom
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  • Nicoletta Bobola,

    1. Core Technology Facility, Faculty of Medical and Human Sciences, and The University of Manchester, Manchester, United Kingdom
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  • Angela Russell,

    1. Department of Pharmacology and Chemistry Research Laboratory, University of Oxford, Oxford, United Kingdom
    2. Department of Chemistry, Chemistry Research Laboratory, University of Oxford, Oxford, United Kingdom
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  • Steve Davies,

    1. Department of Chemistry, Chemistry Research Laboratory, University of Oxford, Oxford, United Kingdom
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  • Rolf Kemler,

    1. Department of Molecular Embryology, Max-Planck Institute of Immunobiology, Freiburg, Germany
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  • Catherine L.R. Merry,

    1. Materials Science Centre, The University of Manchester, Manchester, United Kingdom
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  • Christopher M. Ward

    Corresponding author
    1. Core Technology Facility, Faculty of Medical and Human Sciences, and The University of Manchester, Manchester, United Kingdom
    • Room 2.30d, Core Technology Facility, Faculty of Medical and Human Sciences, The University of Manchester, 46 Grafton Street, M13 9NT, United Kingdom

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    • Telephone: +44 (0)161 275 5182


  • Author contributions: F.S.: conception and design, collection and assembly of data, data analysis and interpretation, manuscript writing, final approval of manuscript; L.M.: conception and design, collection and assembly of data, data analysis and interpretation, manuscript writing; D.E.: provision of study material; S.R.: conception and design, administrative support, collection and assembly of data; A.E.: collection and assembly of data; N.B.: conception and design, data analysis and interpretation, manuscript writing; A.R.: provision of study material; S.D.: provision of study material; R.K.: provision of study material; C.M.: conception and design, collection and assembly of data, data analysis and interpretation, manuscript writing, final approval of manuscript; C.M.W.: conception and design, financial support, provision of study material, collection and assembly of data, data analysis and interpretation, manuscript writing, final approval of manuscript.

  • First published online in STEM CELLS EXPRESS May 28, 2009.

Abstract

We have previously demonstrated that differentiation of embryonic stem (ES) cells is associated with downregulation of cell surface E-cadherin. In this study, we assessed the function of E-cadherin in mouse ES cell pluripotency and differentiation. We show that inhibition of E-cadherin-mediated cell–cell contact in ES cells using gene knockout (Ecad−/−), RNA interference (EcadRNAi), or a transhomodimerization-inhibiting peptide (CHAVC) results in cellular proliferation and maintenance of an undifferentiated phenotype in fetal bovine serum-supplemented medium in the absence of leukemia inhibitory factor (LIF). Re-expression of E-cadherin in Ecad−/−, EcadRNAi, and CHAVC-treated ES cells restores cellular dependence to LIF supplementation. Although reversal of the LIF-independent phenotype in Ecad−/− ES cells is dependent on the β-catenin binding domain of E-cadherin, we show that β-catenin null (βcat−/−) ES cells also remain undifferentiated in the absence of LIF. This suggests that LIF-independent self-renewal of Ecad−/− ES cells is unlikely to be via β-catenin signaling. Exposure of Ecad−/−, EcadRNAi, and CHAVC-treated ES cells to the activin receptor-like kinase inhibitor SB431542 led to differentiation of the cells, which could be prevented by re-expression of E-cadherin. To confirm the role of transforming growth factor β family signaling in the self-renewal of Ecad−/− ES cells, we show that these cells maintain an undifferentiated phenotype when cultured in serum-free medium supplemented with Activin A and Nodal, with fibroblast growth factor 2 required for cellular proliferation. We conclude that transhomodimerization of E-cadherin protein is required for LIF-dependent ES cell self-renewal and that multiple self-renewal signaling networks subsist in ES cells, with activity dependent upon the cellular context. STEM CELLS 2009;27:2069–2080

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