Wnt and Notch Signals Guide Embryonic Stem Cell Differentiation into the Intestinal Lineages§

Authors

  • Soichiro Ogaki,

    1. Stem Cell Biology, Institute of Molecular Embryology and Genetics, Kumamoto University, Honjo, Kumamoto, Japan
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  • Nobuaki Shiraki,

    1. Stem Cell Biology, Institute of Molecular Embryology and Genetics, Kumamoto University, Honjo, Kumamoto, Japan
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  • Kazuhiko Kume,

    1. Stem Cell Biology, Institute of Molecular Embryology and Genetics, Kumamoto University, Honjo, Kumamoto, Japan
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  • Shoen Kume

    Corresponding author
    1. Stem Cell Biology, Institute of Molecular Embryology and Genetics, Kumamoto University, Honjo, Kumamoto, Japan
    2. The Global COE, Kumamoto University, Honjo, Kumamoto, Japan
    • Department of Stem Cell Biology, Institute of Molecular Embryology and Genetics, Kumamoto University, Honjo 2-2-1, Kumamoto 860-0811, Japan
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    • Telephone: +81-96-373-6620; Fax: +81-96-373-6807


  • Author contributions. S.O.: acquisition of data and analysis and interpretation of data; N.S.: study concept and design and acquisition, analysis, and interpretation of data; K.K.: critical revision of the manuscript and administrative and study supervision; S.K.: study concept and design, drafting and revision of the manuscript, approval of the final version of the manuscript, and obtained funding.

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

  • §

    first published online in STEM CELLS EXPRESS February 4, 2013.

Abstract

The studies of differentiation of mouse or human embryonic stem cells (hESCs) into specific cell types of the intestinal cells would provide insights to the understanding of intestinal development and ultimately yield cells for the use in future regenerative medicine. Here, using an in vitro differentiation procedure of pluripotent stem cells into definitive endoderm (DE), inductive signal pathways' guiding differentiation into intestinal cells was investigated. We found that activation of Wnt/β-catenin and inhibition of Notch signaling pathways, by simultaneous application of 6-bromoindirubin-3′-oxime (BIO), a glycogen synthase kinase-3β inhibitor, and N-[(3,5-Difluorophenyl)acetyl]-L-alanyl-2-phenylglycine-1,1-dimethylethyl ester (DAPT), a known γ-secretase inhibitor, efficiently induced intestinal differentiation of ESCs cultured on feeder cell. BIO and DAPT patterned the DE at graded concentrations. Upon prolonged culture on feeder cells, all four intestinal differentiated cell types, the absorptive enterocytes and three types of secretory cells (goblet cells, enteroendocrine cells, and Paneth cells), were efficiently differentiated from mouse and hESC-derived intestinal epithelium cells. Further investigation revealed that in the mouse ESCs, fibroblast growth factor (FGF) and bone morphogenetic protein (BMP) signaling act synergistically with BIO and DAPT to potentiate differentiation into the intestinal epithelium. However, in hESCs, FGF signaling inhibited, and BMP signaling did not affect differentiation into the intestinal epithelium. We concluded that Wnt and Notch signaling function to pattern the anterior-posterior axis of the DE and control intestinal differentiation. STEM Cells 2013;31:1086–1096

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