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

Winzi MK, Hyttel P, Dale JK, Serup P
Stem Cells Dev. 2011 20 (11): 1817-27

PMID: 21351873 · PMCID: PMC3928718 · DOI:10.1089/scd.2011.0042

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.

MeSH Terms (23)

Animals Antigens, Differentiation Benzamides Cell Differentiation Cell Line Cell Proliferation Chick Embryo Dioxoles Embryonic Stem Cells Green Fluorescent Proteins Hepatocyte Nuclear Factor 3-beta Homeodomain Proteins Intercellular Signaling Peptides and Proteins Mice Notochord Organizers, Embryonic Peptide Hormones Pyrroles Receptors, Growth Factor Recombinant Fusion Proteins Signal Transduction Tissue Culture Techniques Transplantation, Heterologous

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