title: FGF4-dependent stem cells derived from rat blastocysts differentiate along the trophoblast lineage.
abstract: Differentiated trophoblast cell lineages arise from trophoblast stem (TS) cells. To date such a stem cell population has only been established in the mouse. The objective of this investigation was to establish TS cell populations from rat blastocysts. Blastocysts were cultured individually on a feeder layer of rat embryonic fibroblasts (REFs) in fibroblast growth factor-4 (FGF4) and heparin supplemented culture medium. Once cell colonies were established REF feeder layers could be replaced with REF conditioned medium. The blastocyst-derived cell lines, in either proliferative or differentiated states, did not express genes indicative of ICM-derived tissues. In the proliferative state the cells expressed established stem cell-associated markers of TS cells. Cells ceased proliferation and differentiated when FGF4, heparin, and REF conditioned medium were removed. Differentiation was characterized by a decline of stem cell-associated marker gene expression, the appearance of large polyploid cells (trophoblast giant cells), and the expression of trophoblast differentiation-associated genes. Collectively, the data indicate that the rat blastocyst-derived cell lines not only possess many features characteristic of mouse TS cells but also possess some distinct properties. These rat TS cell lines represent valuable new in vitro models for analyses of mechanisms controlling TS cell renewal and differentiation.
authors.affiliation: Institute for Reproductive Health and Regenerative Medicine, Department of Pathology, University of Kansas Medical Center, Kansas City, KS 66160, USA.
authors.forename: Kazuo
authors.initials: K
authors.lastname: Asanoma
authors.forename: M A Karim
authors.initials: MA
authors.lastname: Rumi
authors.forename: Lindsey N
authors.initials: LN
authors.lastname: Kent
authors.forename: Damayanti
authors.initials: D
authors.lastname: Chakraborty
authors.forename: Stephen J
authors.initials: SJ
authors.lastname: Renaud
authors.forename: Norio
authors.initials: N
authors.lastname: Wake
authors.forename: Dong-Soo
authors.initials: DS
authors.lastname: Lee
authors.forename: Kaiyu
authors.initials: K
authors.lastname: Kubota
authors.forename: Michael J
authors.initials: MJ
authors.lastname: Soares
mesh_terms: D000818:Animals; D001755:Blastocyst / Q000166:cytology*; D002454:Cell Differentiation; D019070:Cell Lineage; D005260:Female; D051519:Fibroblast Growth Factor 4 / Q000502:physiology*; D008297:Male; D051379:Mice; D010641:Phenotype; D051381:Rats; D017207:Rats, Sprague-Dawley; D013234:Stem Cells / Q000166:cytology*; D014327:Trophoblasts / Q000166:cytology*
chemical_list: C496447:Fgf4 protein, rat; D051519:Fibroblast Growth Factor 4