title: Establishment of trophoblast stem cells under defined culture conditions in mice.
abstract: The inner cell mass (ICM) and trophoblast cell lineages duet early embryonic development in mammals. After implantation, the ICM forms the embryo proper as well as some extraembryonic tissues, whereas the trophoectoderm (TE) exclusively forms the fetal portion of the placenta and the trophoblast giant cells. Although embryonic stem (ES) cells can be derived from ICM in cultures of mouse blastocysts in the presence of LIF and/or combinations of small-molecule chemical compounds, and the undifferentiated pluripotent state can be stably maintained without use of serum and feeder cells, defined culture conditions for derivation and maintenance of undifferentiated trophoblast stem (TS) cells have not been established. Here, we report that addition of FGF2, activin A, XAV939, and Y27632 are necessary and sufficient for derivation of TS cells from both of E3.5 blastocysts and E6.5 early postimplantation extraembryonic ectoderm. Moreover, the undifferentiated TS cell state can be stably maintained in chemically defined culture conditions. Cells derived in this manner expressed TS cell marker genes, including Eomes, Elf5, Cdx2, Klf5, Cdh1, Esrrb, Sox2, and Tcfap2c; differentiated into all trophoblast subtypes (trophoblast giant cells, spongiotrophoblast, and labyrinthine trophoblast) in vitro; and exclusively contributed to trophoblast lineages in chimeric animals. This delineation of minimal requirements for derivation and self-renewal provides a defined platform for precise description and dissection of the molecular state of TS cells.
authors.affiliation: PRESTO, Japan Science and Technology Agency, Kawaguchi, Saitama, Japan; Life Science Experimental Facility, Department of Biotechnology, Faculty of Life and Environmental Sciences, University of Yamanashi, Kofu, Yamanashi, Japan; Laboratory for Pluripotent Stem Cell Studies, Center for Developmental Biology, RIKEN Kobe institute, Kobe Chuo-ku, Hyogo, Japan.
authors.forename: Yasuhide
authors.initials: Y
authors.lastname: Ohinata
authors.affiliation: Laboratory for Pluripotent Stem Cell Studies, Center for Developmental Biology, RIKEN Kobe institute, Kobe Chuo-ku, Hyogo, Japan.
authors.forename: Tomoyuki
authors.initials: T
authors.lastname: Tsukiyama
mesh_terms: D028341:Activins / Q000494:pharmacology; D000577:Amides / Q000494:pharmacology; D000818:Animals; D001755:Blastocyst / Q000187:drug effects / Q000378:metabolism / Q000502:physiology; D002454:Cell Differentiation / Q000187:drug effects / Q000502:physiology; D019070:Cell Lineage / Q000187:drug effects / Q000502:physiology; D003470:Culture Media / Q000378:metabolism*; D004475:Ectoderm / Q000187:drug effects / Q000378:metabolism / Q000502:physiology; D010064:Embryo Implantation / Q000187:drug effects / Q000502:physiology; D047108:Embryonic Development / Q000187:drug effects / Q000502:physiology; D053595:Embryonic Stem Cells / Q000187:drug effects* / Q000378:metabolism / Q000502:physiology*; D005260:Female; D016222:Fibroblast Growth Factor 2 / Q000494:pharmacology; D006575:Heterocyclic Compounds, 3-Ring / Q000494:pharmacology; D006801:Humans; D051379:Mice; D010920:Placenta / Q000378:metabolism / Q000502:physiology; D011247:Pregnancy; D011725:Pyridines / Q000494:pharmacology; D014327:Trophoblasts / Q000187:drug effects* / Q000378:metabolism / Q000502:physiology*
chemical_list: D000577:Amides; D003470:Culture Media; D006575:Heterocyclic Compounds, 3-Ring; D011725:Pyridines; C544261:XAV939; C422405:activin A; D016222:Fibroblast Growth Factor 2; D028341:Activins; C108830:Y 27632