Academic Unit of Reproductive and Developmental Medicine, The University of Sheffield, Level 4, Jessop Wing, Tree Root Walk, Sheffield S10 2SF, UK.
Hum Reprod. 2010 Sep;25(9):2217-28. doi: 10.1093/humrep/deq185. Epub 2010 Jul 23.
Embryo implantation in the uterus involves the trophoblast cells apposing and adhering to, then invading across the epithelium lining of the endometrium. However, ethical concerns regarding experimentation with primary human tissue during this period of life necessitates creation of in vitro models for understanding the basic mechanisms involved. Toll-like receptors (TLRs) play a crucial role in defence against pathogens invading the female reproductive tract. The objective of this study is to establish and optimize an in vitro model for studying human endometrial embryonic interactions and to understand the effect of TLR5 stimulation on the attachment of trophoblast cells to endometrial cells.
By using a human telomerase immortalized endometrial epithelial cell line (hTERT-EECs) and choriocarcinoma human trophoblast cells (JAr cells), an in vitro assay of human implantation was established. In order to investigate the impact of TLR5 stimulation on attachment in this assay, bacterial flagellin was applied to the endometrial and trophoblast cells. In order to block TLR5 in the endometrial and trophoblast cells, TLR5 function-blocking antibody was applied to the cells prior to flagellin treatment.
The results demonstrated that JAr spheroids attached to hTERT-EECs in a time and concentration-dependent manner. Our results also demonstrated that treatment of endometrial cells with flagellin, suppressed the attachment of JAr spheres to the endometrial cells. Application of TLR5 function-blocking antibody significantly restored the attachment of JAr spheres to the endometrium.
These data suggest a novel mechanism by which the presence of intrauterine infection through TLR5 activation may result in implantation failure. These data may provide a new opportunity in the management of infertility cases.
胚胎在子宫内着床涉及滋养层细胞附着和黏附，然后穿过子宫内膜上皮层进行侵袭。然而，在生命的这个阶段，出于对用原代人组织进行实验的伦理关注，需要创建体外模型来理解所涉及的基本机制。Toll 样受体（TLR）在防御女性生殖道入侵病原体方面发挥着至关重要的作用。本研究的目的是建立和优化体外模型，以研究人类子宫内膜与胚胎的相互作用，并了解 TLR5 刺激对滋养层细胞附着在内膜细胞上的影响。
通过使用人端粒酶永生化子宫内膜上皮细胞系（hTERT-EECs）和绒毛膜癌细胞系（JAr 细胞），建立了人类植入的体外模型。为了研究 TLR5 刺激对该模型中附着的影响，将细菌鞭毛蛋白应用于子宫内膜和滋养层细胞。为了阻断 TLR5 在子宫内膜和滋养层细胞中的作用，在鞭毛蛋白处理之前将 TLR5 功能阻断抗体应用于细胞。
结果表明，JAr 球体以时间和浓度依赖的方式附着于 hTERT-EECs。我们的结果还表明，用鞭毛蛋白处理子宫内膜细胞会抑制 JAr 球体附着到子宫内膜细胞上。应用 TLR5 功能阻断抗体可显著恢复 JAr 球体与子宫内膜的附着。
这些数据表明，通过 TLR5 激活宫内感染的存在可能导致着床失败的一种新机制。这些数据可能为管理不孕病例提供新的机会。
