Department of Obstetrics and Gynecology, The Jones Institute for Reproductive Medicine, Eastern Virginia Medical School, Norfolk, Virginia, USA.
Tissue Eng Part C Methods. 2013 Sep;19(9):676-87. doi: 10.1089/ten.TEC.2012.0616. Epub 2013 Feb 19.
Human embryo implantation involves a complex network of molecular signaling that is modulated by endocrine and paracrine pathways. Here, we performed studies using a unique and recently developed three-dimensional (3D) implantation model, characterized by an endometrium-like 3D culture system and Jar cell-derived spheroids mimicking the embryo/trophoblast. The aims were to investigate the effects of 17β estradiol (E2) and medroxyprogesterone acetate (MPA) on (1) the interaction between epithelial and stromal cells, and (2) the attachment and invasion of trophoblast cells. We observed that epithelial and stromal cells in the 3D culture were ERα⁺, ERβ⁺, and PR⁺. Decidualization was confirmed by enhanced prolactin gene expression on day 7 of E2 plus MPA treatment. An effect of epithelial cells on the decidualization of stromal cells was indicated by significantly higher levels of prolactin mRNA expression in the 3D culture compared to stromal cells grown within the fibrin-agarose gel matrix. On the other hand, the relative gene expressions of E-cadherin and IL-1β in epithelial cells of the 3D culture under decidualization conditions significantly differed from those in epithelial cells grown over the fibrin-agarose gel matrix without stromal cells, pointing to regulation of epithelial cells by the stroma. The attachment rate of Jar spheroids to the 3D was significantly increased by E2 plus MPA treatment. Analyses of Z-stack confocal and stained optic microscopic images demonstrated that Jar spheroids breached the epithelial cell monolayer, invaded, and were embedded into the 3D matrix in response to decidualization signals. In summary, the newly bioengineered system provides a unique model for studying interactions between the different endometrial cell compartments, via soluble-paracrine signals as well as cell-to-cell interactions, and is a useful tool to study early embryonic implantation events.
人类胚胎着床涉及一个复杂的分子信号网络，该网络受内分泌和旁分泌途径的调节。在这里，我们使用了一种独特的、最近开发的三维（3D）着床模型进行研究，该模型的特点是具有类似于子宫内膜的 3D 培养系统和 Jar 细胞衍生的球体，可模拟胚胎/滋养层。目的是研究 17β 雌二醇（E2）和醋酸甲羟孕酮（MPA）对（1）上皮细胞和基质细胞之间的相互作用，以及（2）滋养层细胞的附着和侵袭的影响。我们观察到，3D 培养中的上皮细胞和基质细胞均为 ERα⁺、ERβ⁺和 PR⁺。E2 加 MPA 处理第 7 天催乳素基因表达增强证实了蜕膜化。上皮细胞对基质细胞蜕膜化的影响表明，与在纤维蛋白-琼脂糖凝胶基质中生长的基质细胞相比，3D 培养中的催乳素 mRNA 表达水平显著更高。另一方面，在蜕膜化条件下，3D 培养中的上皮细胞的 E-钙粘蛋白和 IL-1β的相对基因表达与在没有基质细胞的纤维蛋白-琼脂糖凝胶基质中生长的上皮细胞明显不同，这表明上皮细胞受基质的调节。Jar 球体在 3D 上的附着率在 E2 加 MPA 处理后显著增加。Z 堆叠共聚焦和染色光学显微镜图像分析表明，Jar 球体突破上皮细胞单层，侵袭并嵌入 3D 基质，以响应蜕膜化信号。总之，新的生物工程系统为研究不同子宫内膜细胞区室之间的相互作用提供了一个独特的模型，通过可溶性旁分泌信号以及细胞间相互作用，并为研究早期胚胎着床事件提供了有用的工具。
