Department of Obstetrics and Gynaecology, Monash University, Melbourne, Victoria, Australia; Centre for Reproductive Health, Hudson Institute of Medical Research, Clayton, Victoria, Australia.
Molecular Proteomics Laboratory, Baker Heart and Diabetes Institute, Melbourne, Victoria, Australia.
Mol Hum Reprod. 2020 Jul 1;26(7):510-520. doi: 10.1093/molehr/gaaa034.
A successful pregnancy requires a synchronous dialogue between endometrium and embryo within the endometrial milieu. The aim of this study was to assess the role in the implantation of mediators in the endometrial milieu. Total secretome (TS), soluble secretome (SS) and small extracellular vesicles (containing exosomes) were generated from hormonally primed human endometrial epithelial cell culture medium. Human trophectoderm stem cell-derived spheroids were cultured with TS, SS or exosomes (30 µg/ml) on hormonally primed epithelial cells, with exosomes significantly increasing cell adhesion and outgrowth. Furthermore, F1 mouse 2-cell embryos were cultured in groups for 48 h followed by culture with each secretome fraction (30 µg/ml) for 48 h. Blastocyst cell number and hatching were quantified. In addition, blastocysts were further cultured on a fibronectin matrix for 72 h or transferred to recipient mice (with corresponding secretomes) with embryo implantation assessed after 6 days. Exosomes significantly increased total cell number in mouse embryos and complete hatching from zona pellucida, with both exosomes and SS significantly enhancing mouse embryo outgrowth. Importantly, exosomes increased the embryo implantation rate in comparison to other secretome fractions (normalized based on treatment amount) from the endometrial epithelia. These data indicate that endometrial epithelial exosomes support embryo growth, development and implantation while the SS has selective involvement specifically on mouse embryo outgrowth. This finding provides new insights into the molecular differences of endometrial secretome components in implantation and early embryo development and may implicate endometrial exosomes in the pathophysiology of implantation failure in infertility.
成功的妊娠需要子宫内膜和胚胎在子宫内膜环境中进行同步对话。本研究旨在评估子宫内膜环境中介质在植入中的作用。从激素预处理的人子宫内膜上皮细胞培养基中生成总分泌组（TS）、可溶分泌组（SS）和小细胞外囊泡（包含外泌体）。将人滋养外胚层干细胞衍生的球体与 TS、SS 或外泌体（30μg/ml）在激素预处理的上皮细胞上共培养，外泌体显著增加细胞黏附和生长。此外，将 F1 小鼠 2 细胞胚胎分组培养 48 小时，然后用每种分泌组（30μg/ml）培养 48 小时。量化胚胎囊胚细胞数量和孵化情况。此外，将囊胚进一步在纤维连接蛋白基质上培养 72 小时，或转移到接受者小鼠（具有相应的分泌组）中，在 6 天后评估胚胎植入情况。外泌体显著增加了小鼠胚胎的总细胞数量，并从透明带中完全孵化，外泌体和 SS 均显著增强了小鼠胚胎的生长。重要的是，与其他分泌组（基于处理量进行归一化）相比，外泌体增加了胚胎的植入率。这些数据表明，子宫内膜上皮细胞外泌体支持胚胎生长、发育和植入，而 SS 则特别参与小鼠胚胎的生长。这一发现为植入和早期胚胎发育中子宫内膜分泌组成分的分子差异提供了新的见解，并可能暗示子宫内膜外泌体在不孕中植入失败的病理生理学中的作用。
