title: Isolation and characterization of node/notochord-like cells from mouse embryonic stem cells.
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.
authors.affiliation: Department of Stem Cell Biology, Hagedorn Research Institute, Gentofte, Denmark.
authors.forename: Maria K
authors.initials: MK
authors.lastname: Winzi
authors.forename: Poul
authors.initials: P
authors.lastname: Hyttel
authors.forename: Jacqueline Kim
authors.initials: JK
authors.lastname: Dale
authors.forename: Palle
authors.initials: P
authors.lastname: Serup
mesh_terms: D000818:Animals; D000943:Antigens, Differentiation / Q000378:metabolism; D001549:Benzamides / Q000494:pharmacology; D002454:Cell Differentiation / Q000187:drug effects; D002460:Cell Line; D049109:Cell Proliferation; D002642:Chick Embryo; D004149:Dioxoles / Q000494:pharmacology; D053595:Embryonic Stem Cells / Q000166:cytology* / Q000187:drug effects / Q000378:metabolism / Q000637:transplantation; D049452:Green Fluorescent Proteins / Q000096:biosynthesis; D051542:Hepatocyte Nuclear Factor 3-beta / Q000494:pharmacology / Q000502:physiology; D018398:Homeodomain Proteins / Q000096:biosynthesis; D036341:Intercellular Signaling Peptides and Proteins / Q000494:pharmacology / Q000502:physiology; D051379:Mice; D009672:Notochord / Q000166:cytology*; D020897:Organizers, Embryonic / Q000166:cytology* / Q000378:metabolism; D036361:Peptide Hormones / Q000494:pharmacology / Q000502:physiology; D011758:Pyrroles / Q000494:pharmacology; D017978:Receptors, Growth Factor / Q000819:agonists / Q000037:antagonists & inhibitors; D011993:Recombinant Fusion Proteins / Q000096:biosynthesis; D015398:Signal Transduction; D046509:Tissue Culture Techniques; D014183:Transplantation, Heterologous
chemical_list: C459179:4-(5-benzo(1,3)dioxol-5-yl-4-pyridin-2-yl-1H-imidazol-2-yl)benzamide; D000943:Antigens, Differentiation; D001549:Benzamides; D004149:Dioxoles; D018398:Homeodomain Proteins; D036341:Intercellular Signaling Peptides and Proteins; C489656:Not protein, mouse; D036361:Peptide Hormones; D011758:Pyrroles; D017978:Receptors, Growth Factor; D011993:Recombinant Fusion Proteins; C105686:SU 5402; D051542:Hepatocyte Nuclear Factor 3-beta; D049452:Green Fluorescent Proteins