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Establishment of Trophoblast Stem Cells under Defined Culture Conditions in Mice

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 gia...

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Autores principales: Ohinata, Yasuhide, Tsukiyama, Tomoyuki
Formato: Online Artículo Texto
Lenguaje:English
Publicado: Public Library of Science 2014
Materias:
Acceso en línea:https://www.ncbi.nlm.nih.gov/pmc/articles/PMC4159327/
https://www.ncbi.nlm.nih.gov/pubmed/25203285
http://dx.doi.org/10.1371/journal.pone.0107308
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author Ohinata, Yasuhide
Tsukiyama, Tomoyuki
author_facet Ohinata, Yasuhide
Tsukiyama, Tomoyuki
author_sort Ohinata, Yasuhide
collection PubMed
description 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.
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spelling pubmed-41593272014-09-12 Establishment of Trophoblast Stem Cells under Defined Culture Conditions in Mice Ohinata, Yasuhide Tsukiyama, Tomoyuki PLoS One Research Article 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. Public Library of Science 2014-09-09 /pmc/articles/PMC4159327/ /pubmed/25203285 http://dx.doi.org/10.1371/journal.pone.0107308 Text en © 2014 Ohinata, Tsukiyama http://creativecommons.org/licenses/by/4.0/ This is an open-access article distributed under the terms of the Creative Commons Attribution License, which permits unrestricted use, distribution, and reproduction in any medium, provided the original author and source are properly credited.
spellingShingle Research Article
Ohinata, Yasuhide
Tsukiyama, Tomoyuki
Establishment of Trophoblast Stem Cells under Defined Culture Conditions in Mice
title Establishment of Trophoblast Stem Cells under Defined Culture Conditions in Mice
title_full Establishment of Trophoblast Stem Cells under Defined Culture Conditions in Mice
title_fullStr Establishment of Trophoblast Stem Cells under Defined Culture Conditions in Mice
title_full_unstemmed Establishment of Trophoblast Stem Cells under Defined Culture Conditions in Mice
title_short Establishment of Trophoblast Stem Cells under Defined Culture Conditions in Mice
title_sort establishment of trophoblast stem cells under defined culture conditions in mice
topic Research Article
url https://www.ncbi.nlm.nih.gov/pmc/articles/PMC4159327/
https://www.ncbi.nlm.nih.gov/pubmed/25203285
http://dx.doi.org/10.1371/journal.pone.0107308
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