An in-vitro model for stromal invasion during implantation of the human blastocyst
An in-vitro model for stromal invasion during implantation of the human blastocyst
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DOI：
10.1093/humrep/deg072
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发表时间：
2003-02-01
影响因子：
6.1
通讯作者：
Mardon, H
中科院分区：
文献类型：
--
作者：
Carver, J;Martin, K;Mardon, H
背景：植入失败可能是不孕不育的主要原因。在小鼠身上的研究已经确定了一些参与植入的分子，但在人类体内的植入机制仍不清楚，这主要是因为缺乏提供功能信息的人体植入模型。方法：将孵化的人胚泡与人子宫内膜基质细胞单层共培养。对植入胚泡进行延时照相，细胞角蛋白和肌动蛋白免疫组织化学染色，测定培养上清液中hCG的分泌。结果：胚泡附着于基质细胞层，并植入基质细胞层。滋养层细胞向基质细胞层生长和侵袭主要发生在两个相对的两极，其方向与基质细胞成纤维细胞的方向一致。高分辨率图像分析显示滋养层细胞完全穿透基质细胞层。整个植入部位的免疫染色显示出靠近滋养细胞基面的独特的肌动蛋白和细胞角蛋白阳性锚定结构。与塑料培养的胚泡相比，植入基质细胞的胚泡分泌更高水平的hCG。结论：为研究人类胚胎植入子宫内膜间质的机制建立了一个可靠的模型。
BACKGROUND: Implantation failure is likely to be a major cause of infertility. Studies in mice have identified a number of molecules that are involved in implantation, but the mechanisms of implantation in the human remain unclear, largely due to the lack of models for implantation in the human that provide functional information. METHODS: Human hatched blastocysts were co-cultured with human endometrial stromal cell monolayers. Time-lapse photography of implanting blastocysts, immunostaining for cytokeratin and actin, and measurement of hCG secreted into the culture supernatants were performed. RESULTS: Blastocysts attached to and implanted into the stromal cell layer. Trophoblast outgrowth onto, and invasion into, the stromal cell layer occurred largely at two opposite poles, the orientation of which was aligned to that of the stromal cell fibroblasts. High-resolution image analysis demonstrated that the trophoblast completely penetrated the stromal cell layer. Immunostaining of whole-mounts of implantation sites revealed distinctive actin and cytokeratin-positive anchoring structures adjacent to the basal surface of the trophoblast. Blastocysts implanting into stromal cells secreted higher levels of hCG compared with those cultured on plastic. CONCLUSIONS: A robust model for the study of mechanisms of implantation of the human embryo into the endometrial stroma has been established.