Establishment and controlled differentiation of neural crest stem cell lines using conditional transgenesis

Authors

  • Jochen Maurer,

    1. Department of Developmental Pathology, Institute for Pathology, University of Bonn Medical School, Sigmund-Freud-Strasse 25 53127 Bonn, Germany
      Tel: +49 228 287 6342
      Fax: +49 228 287 9757
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  • Sebastian Fuchs,

    1. Institute of Cell Biology, Swiss Federal Institute of Technology, ETH-Hönggerberg HPM E38, CH-8093 Zürich, Switzerland
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  • Richard Jäger,

    1. Department of Developmental Pathology, Institute for Pathology, University of Bonn Medical School, Sigmund-Freud-Strasse 25 53127 Bonn, Germany
      Tel: +49 228 287 6342
      Fax: +49 228 287 9757
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  • Bodo Kurz,

    1. Anatomisches Institut, Christian-Albrechts-Universität zu Kiel, Olshausenstr 40 24098 Kiel, Germany
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  • Lukas Sommer,

    1. Institute of Cell Biology, Swiss Federal Institute of Technology, ETH-Hönggerberg HPM E38, CH-8093 Zürich, Switzerland
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  • Hubert Schorle

    Corresponding author
    1. Department of Developmental Pathology, Institute for Pathology, University of Bonn Medical School, Sigmund-Freud-Strasse 25 53127 Bonn, Germany
      Tel: +49 228 287 6342
      Fax: +49 228 287 9757
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✉ E-mail: Hubert.Schorle@ukb.uni-bonn.de

Abstract

Abstract Murine neural crest stem cells (NCSCs) are a multipotent transient population of stem cells. After being formed during early embryogenesis as a consequence of neurulation at the apical neural fold, the cells rapidly disperse throughout the embryo, migrating along specific pathways and differentiating into a wide variety of cell types. In vitro the multipotency is lost rapidly, making it difficult to study differentiation potential as well as cell fate decisions. Using a transgenic mouse line, allowing for spatio-temporal control of the transforming c-myc oncogene, we derived a cell line (JoMa1), which expressed NCSC markers in a transgene-activity dependent manner. JoMa1 cells express early NCSC markers and can be instructed to differentiate into neurons, glia, smooth muscle cells, melanocytes, and also chondrocytes. A cell-line, clonally derived from JoMa1 culture, termed JoMa1.3 showed identical behavior and was studied in more detail. This system therefore represents a powerful tool to study NCSC biology and signaling pathways. We observed that when proliferative and differentiation stimuli were given, enhanced cell death could be detected, suggesting that the two signals are incompatible in the cellular context. However, the cells regain their differentiation potential after inactivation of c-MycERT. In summary, we have established a system, which allows for the biochemical analysis of the molecular pathways governing NCSC biology. In addition, we should be able to obtain NCSC lines from crossing the c-MycERT mice with mice harboring mutations affecting neural crest development enabling further insight into genetic pathways controlling neural crest differentiation.

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