Isolation and characterization of node/notochord-likecCells from mouse embryonic stem cells

Maria K. Winzi, Poul Hyttel, Jacqueline Kim Dale, Palle Serup

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    13 Citations (Scopus)

    Abstract

    The homeobox gene Noto is expressed in the node and its derivative the notochord. Here we use a targeted Noto-GFP reporter to isolate and characterize node/notochord-like cells derived from mouse embryonic stem cells. We find very few Noto-expressing cells after spontaneous differentiation. However, the number of Noto-expressing cells was increased when using Activin A to induce a Foxa2- and Brachyury-expressing progenitor population, whose further differentiation into Noto-expressing cells was improved by simultaneous inhibition of BMP, Wnt, and retinoic acid signaling. Noto-GFP(+) cells expressed the node/notochord markers Noto, Foxa2, Shh, Noggin, Chordin, Foxj1, and Brachyury; showed a vacuolarization characteristic of notochord cells; and can integrate into midline structures when grafted into Hensen's node of gastrulating chicken embryos. The ability to generate node/notochord-like cells in vitro will aid the biochemical characterization of these developmentally important structures.

    Original languageEnglish
    Pages (from-to)1817-1827
    Number of pages11
    JournalStem Cells and Development
    Volume20
    Issue number11
    Early online date6 Apr 2011
    DOIs
    Publication statusPublished - Nov 2011

    Keywords

    • Fibroblast growth factor
    • Floor plate
    • Sonic hedgehog
    • Notochord formation
    • Pancreas development
    • Expression patterns
    • Definitive endoderm
    • Retinoic acid
    • Nodal flow
    • Gene

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