Project description:Trophectoderm-specific expression of Angiomotin (AMOT) in pre-implantation embryos followed by its unique expression in the post-implantation ectoplacental cone that harbors the trophoblast stem cell niche prompted our investigation on the function of AMOT in trophoblast cells. Using the in vitro trophoblast stem cell culture model, we established differentiation dependent up-regulation of AMOT expression in trophoblast cells. To understand the function of AMOT in trophoblast cells mass spectrometry-based proteomic analysis was employed to identify the AMOT interactome within the trophoblast proteome. This approach utilized immunoprecipitation of endogenous AMOT followed by fractionation on SDS-PAGE and subsequently subjecting the tryptic digested excised gel bands to mass spectrometry.
Project description:As medical abortion has established itself as the significant method of terminating pregnancies, many studies in recent years has focused on Mifepristone, a principal agent used in this field. It can be found that much effort has been devoted to the research on the impact of Mifepristone on the decidua and embryo implantation process in the mechanism of abortion and contraception; the mechanism of its impact on trophoblast cells, however, still remains unclear. Therefore, the present study aims to fill the gap by investigating the effects of Mifepristone on villous trophoblasts in medical abortion patients. We have found that Mifepristone affects trophoblast cell proliferation and invasion through the progesterone receptor. We established PGR-knockdown trophoblast stem cells and collected these cells for comprehensive RNA-seq analysis.
Project description:The placenta serves as a critical interface between the mother and fetus, with placental dysfunction being a major contributor to various pregnancy complications. Successful placental formation relies on the robust expansion of trophoblast stem cells, followed by precisely regulated lineage specification. Trophoblast stem cells (TSCs) have emerged as a valuable model for investigating placentation and related disorders. However, despite their ability to be maintained over multiple passages without losing stemness, conventional TSCs are characterized by inherent heterogeneity and a spontaneous tendency to differentiate autonomously. In this study, we established a novel type of mouse trophoblast stem cell. These cells form dome-shaped colonies, morphologically distinct from tight epithelial classic TSC colonies with a raised edge, and are therefore termed dome-shaped TSCs (dTSCs). The established dTSCs maintain the dome-shaped morphology, express canonical TSC markers, exhibit reduced cellular heterogeneity, display a relatively low propensity for spontaneous differentiation under stem cell maintenance conditions, retain multipotency for differentiating into specialized trophoblast lineages, and are capable of integrating into E12.5 placentas in chimeric assays. Meanwhile, dTSCs are amenable to gene manipulations, providing a practical means for studying functional genomics in the trophoblast lineages. Collectively, these findings establish dTSCs as a promising and refined model for studying trophoblast development.
