Abstract
The placenta forms a maternal-fetal junction that supports many physiological functions such as the supply of nutrition and exchange of gases and wastes. Establishing an in vitro culture model of human and non-human primate trophoblast stem/progenitor cells is important for investigating the process of early placental development and trophoblast differentiation. In this study, we have established five trophoblast stem cell (TSC) lines from cynomolgus monkey blastocysts, named macTSC #1-5. Fibroblast growth factor 4 (FGF4) enhanced proliferation of macTSCs, while other exogenous factors were not required to maintain their undifferentiated state. macTSCs showed a trophoblastic gene expression profile and trophoblast-like DNA methylation status and also exhibited differentiation capacity towards invasive trophoblast cells and multinucleated syncytia. In a xenogeneic chimera assay, these stem cells contributed to trophectoderm (TE) development in the chimeric blastocysts. macTSC are the first primate trophoblast cell lines whose proliferation is promoted by FGF4. These cell lines provide a valuable in vitro culture model to analyze the similarities and differences in placental development between human and non-human primates.
Highlights
The placenta forms a maternal-fetal junction that supports many physiological functions such as the supply of nutrition and exchange of gases and wastes
We attempted to establish trophoblastic cell lines from cynomolgus monkey blastocysts by adopting the conditions used for the establishment and maintenance of mouse trophoblast stem cell (TSC), with slight modifications
For the maintenance of mouse TSCs, mouse embryonic fibroblast cells (MEF) can be replaced by the MEF-conditioned medium (MEF-CM) or by activin A20,21
Summary
The placenta forms a maternal-fetal junction that supports many physiological functions such as the supply of nutrition and exchange of gases and wastes. Establishing an in vitro culture model of human and non-human primate trophoblast stem/progenitor cells is important for investigating the process of early placental development and trophoblast differentiation. Previous reports show the establishment of TSC lines from rhesus monkey blastocysts[18] These cell lines were capable of differentiating into both invasive cells and multinucleated cells; they expressed the ICM/ESC marker, OCT4, and the marker for STBs, chorionic gonadotropin (CG)[18,19], without induction of differentiation, making it uncertain if these cells are genuine TSCs. the establishment of trophoblastic stem/progenitor cell lines from a non-human primate, whose placental structure is similar to that of humans, remains an important issue.
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