AN IN VITRO THREE DIMENSIONAL MODEL SIMULATING PRIMATE EMBRYO IMPLANTATION

本子项目是利用由NIH/NCRR资助的中心赠款提供的资源的众多研究子项目之一。子项目和研究者（PI）可能已经从另一个NIH来源获得了主要资金，因此可以在其他CRISP条目中表示。列出的机构是中心的，不一定是研究者的机构。建立非人类灵长类动物胚胎的三维模型，模拟胚胎着床和着床后的发育。利用体外受精过程中产生的恒河猴（猕猴）胚胎产生囊胚。将扩增囊胚（n= 42）包埋于三维基质环境中，并在水牛大鼠肝细胞条件培养基中培养。胚泡包埋后第5 ~ 6天，胚泡植入体表面出现细胞样突起，胚泡开始孵化，细胞生长和囊性形成。此外，在三维环境中还观察到类似于活体胎盘结构的分支状突起。免疫分析显示CG和黄体酮分泌持续升高。对基质细胞外植体胚胎的形态学分析表明，基质细胞有广泛的滋养层分化和迁移，金属蛋白酶对基质细胞环境的降解，以及vimentin表达证实的非滋养层细胞系的分化。这可能是研究早期谱系决定和灵长类胚胎形态发生的一个更容易处理的系统。这项研究使用了WNPRC动物服务和化验服务。

This subproject is one of many research subprojects utilizing the resources provided by a Center grant funded by NIH/NCRR. The subproject and investigator (PI) may have received primary funding from another NIH source, and thus could be represented in other CRISP entries. The institution listed is for the Center, which is not necessarily the institution for the investigator. To develop a three-dimensional model with non-human primate embryos to simulate implantation and post- implantation development. Rhesus monkey (macaca mulatta) embryos generated from IVF procedure were used for producing blastocysts. Expanded blastocysts (n= 42) were embedded into a three dimensional environment of Matrigel and cultured in Buffalo rat liver cell-conditioned medium. Hatching of the blastocysts occurred and cell outgrowths and cystic formation were observed beginning at day 5 to 6 post embedding following a rapid development of cellular-like protrusions on the surface of blastocyst implant. In addition, branch-like projections grown into the three- dimensional environment similar to in vivo placental structures were observed. Immunoassay indicated consistent elevations in CG and progesterone secretion. Morphological analysis of embryos in Matrigel explants revealed extensive trophoblast differentiation and migration, degradation of the Matrigel environment by metalloproteinases, and differentiation of non-trophoblast lineages as evidenced by vimentin expresson. This may be a more tractable system for studying early lineage decisions and morphogenesis in the primate embryo. This research used WNPRC Animal Services and Assay Services.