AN IN VITRO THREE DIMENSIONAL MODEL SIMULATING PRIMATE EMBRYO IMPLANTATION

这个子项目是利用由NIH/NCRR资助的中心拨款提供的资源的许多研究子项目之一。子项目和调查员(PI)可能从另一个NIH来源获得了主要资金，因此可能会出现在其他CRISE条目中。列出的机构是针对中心的，而不一定是针对调查员的机构。建立非人灵长类胚胎的三维模型，模拟着床及着床后发育。将体外受精过程中产生的猕猴(Macaca Mulatta)胚胎用于生产囊胚。将42个扩张的囊胚植入Matrigel的三维环境中，并在水牛肝细胞条件培养液中培养。随着胚泡表面细胞样突起的迅速发展，胚泡在植入后第5~6天开始孵化，并观察到细胞突起和囊泡的形成。此外，观察到类似于体内胎盘结构的树枝状突起生长到三维环境中。免疫分析显示CG和孕酮分泌持续升高。Matrigel外植体胚胎的形态分析显示，滋养层细胞广泛分化和迁移，金属蛋白酶降解Matrigel环境，波形蛋白表达证明非滋养层细胞系分化。对于研究灵长类胚胎的早期谱系决定和形态发生，这可能是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.