Reproductive Medicine Center, Tongji Hospital, Tongji Medical College, Huazhong University of Science and Technology, Jiefang Avenue 1095#, Wuhan, 430030, People's Republic of China.
J Assist Reprod Genet. 2021 Apr;38(4):825-833. doi: 10.1007/s10815-021-02093-5. Epub 2021 Feb 1.
Endometrial extracellular vesicles are essential in regulating trophoblasts' function. This study aims to investigate whether endometrial extracellular vesicles (EVs) from recurrent implantation failure (RIF) patients inhibit the proliferation, invasion, and migration of HTR8/SVneo cells.
Eighteen RIF patients and thirteen fertile women were recruited for endometria collection. Endometrial cells isolated from the endometria were cultured and modulated by hormones, and the conditioned medium was used for EV isolation. EVs secreted by the endometrial cells of RIF patients (RIF-EVs) or fertile women (FER-EVs) were determined by Western blotting, nanoparticle tracking analysis, and transmission electron microscopy. Fluorescence-labeled EVs were used to visualize internalization by HTR8/SVneo cells. RIF-EVs and FER-EVs were co-cultured with HTR8/SVneo cells. Cell Counting Kit-8, transwell invasion, and wound closure assays were performed to determine cellular proliferation, invasion, and migration, respectively, in different treatments.
RIF-EVs and FER-EVs were bilayer membrane vesicles, ranging from 100 to 150 nm in size, that expressed the classic EV markers Alix and CD9. RIF-EVs and FER-EVs were internalized by HTR8/SVneo cells within 2 h. The proliferation rate in the FER-EV group was significantly higher than that in the RIF-EV group at 20 μg/mL. Moreover, the invasion and migration capacity of trophoblast cells were decreased in the RIF-EV group relative to the FER-EV group at 20 μg/mL.
Endometrial EVs from RIF patients inhibited the functions of trophoblasts by decreasing their proliferation, migration, and invasive capacity. Such dysregulations induced by RIF-EVs may provide novel insights for better understanding the pathogenesis of implantation failure.
子宫内膜细胞外囊泡在调节滋养细胞功能方面至关重要。本研究旨在探讨复发性植入失败（RIF）患者的子宫内膜细胞外囊泡（EVs）是否抑制 HTR8/SVneo 细胞的增殖、侵袭和迁移。
招募 18 名 RIF 患者和 13 名生育能力正常的妇女进行子宫内膜采集。从子宫内膜中分离出的子宫内膜细胞经激素培养和调节，并用条件培养基分离 EV。通过 Western blot、纳米颗粒跟踪分析和透射电子显微镜确定 RIF 患者（RIF-EVs）或生育能力正常的妇女（FER-EVs）子宫内膜细胞分泌的 EVs。荧光标记的 EV 用于可视化 HTR8/SVneo 细胞的内化。将 RIF-EVs 和 FER-EVs 与 HTR8/SVneo 细胞共培养。通过细胞计数试剂盒-8、Transwell 侵袭和划痕愈合实验分别检测不同处理方式下细胞的增殖、侵袭和迁移。
RIF-EVs 和 FER-EVs 是双层膜囊泡，大小在 100 至 150nm 之间，表达经典的 EV 标志物 Alix 和 CD9。RIF-EVs 和 FER-EVs 在 2 小时内被 HTR8/SVneo 细胞内化。在 20μg/mL 时，FER-EV 组的增殖率明显高于 RIF-EV 组。此外，在 20μg/mL 时，RIF-EV 组滋养细胞的侵袭和迁移能力较 FER-EV 组降低。
RIF 患者的子宫内膜 EV 通过降低滋养细胞的增殖、迁移和侵袭能力来抑制其功能。RIF-EVs 引起的这种失调可能为更好地理解植入失败的发病机制提供新的见解。
