Isolation and Characterization of Node/Notochord-like Cells from Mouse Embryonic Stem Cells

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  • Maria Karin Winzi, Danmark
  • Poul Maddox-Hyttel
  • ,
  • J Kim Dale
  • ,
  • Palle Serup
  • Molekylærbiologisk Institut
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 ES 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 vacuolarisation 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.
OriginalsprogEngelsk
TidsskriftStem Cells and Development
ISSN1547-3287
DOI
StatusUdgivet - 25 feb. 2011

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