OBJECTIVE To investigate whether endometrial extracellular vesicles (EVs) from patients with recurrent implantation failure (RIF) attenuate the growth and invasion of embryos. DESIGN In vitro experimental study. SETTING University-affiliated hospital. PATIENT(S) Ten RIF patients and seven fertile women. INTERVENTIONS(S) Endometrial cells isolated from endometrial tissues obtained from patients with RIF and fertile women were cultured and modulated in vitro via hormones. Conditioned medium was collected for EV isolation. MAIN OUTCOME MEASURE(S) EVs secreted by endometrial cells of patients with RIF (RIF-EVs) or fertile women (FER-EVs) were characterized with the use of Western blotting, nanoparticle tracking analysis, and transmission electron microscopy. EVs from the two groups were co-cultured with 2-cell murine embryos. Fluorescence-labeled EVs were used to visualize internalization by embryos. Following co-culture, blastocyst and hatching rates were calculated. Blastocysts were stained with diamidino-2-phenylindole to count the total cell number, and the hatched embryos were used to test invasion capacity. RESULT(S) RIF-EVs and FER-EVs are bilayered vesicles ∼100 nm in size and enriched with TSG101, Alix, and CD9. EVs were internalized within 12 hours. The blastocyst rates in the RIF-EV groups were significantly decreased compared with the FER-EV groups at 5, 10, and 20 μg/mL. The hatching rates and total cell numbers of blastocysts also were decreased significantly in the RIF-EV groups compared with the FER-EV groups at 10 and 20 μg/mL. Moreover, the invasion capacity of hatched embryos decreased significantly in the RIF-EV group. CONCLUSION(S) Endometrial EVs from patients with RIF attenuate the development and invasion of embryos.

中文翻译：

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复发性着床失败女性的子宫内膜细胞外囊泡减弱胚​​胎的生长和侵袭

目的 研究复发性着床失败（RIF）患者的子宫内膜细胞外囊泡（EVs）是否减弱胚胎的生长和侵袭。设计 体外实验研究。设置 大学附属医院。患者 10 名 RIF 患者和 7 名可育妇女。干预措施 从 RIF 患者和生育妇女的子宫内膜组织中分离出的子宫内膜细胞在体外通过激素进行培养和调节。收集条件培养基用于 EV 隔离。主要结果测量 RIF (RIF-EVs) 或可育妇女 (FER-EVs) 的子宫内膜细胞分泌的 EVs 使用蛋白质印迹、纳米粒子跟踪分析和透射电子显微镜进行表征。来自两组的 EV 与 2 细胞鼠胚胎共培养。荧光标记的 EV 用于可视化胚胎的内化。共培养后，计算囊胚和孵化率。囊胚用二脒基-2-苯基吲哚染色计算细胞总数，孵化胚胎用于检测侵袭能力。结果 RIF-EVs 和 FER-EVs 是双层囊泡，大小约为 100 nm，富含 TSG101、Alix 和 CD9。EVs 在 12 小时内被内化。与 FER-EV 组相比，RIF-EV 组的囊胚率在 5、10 和 20 μg/mL 时显着降低。与 FER-EV 组相比，RIF-EV 组的囊胚孵化率和总细胞数在 10 和 20 μg/mL 时也显着降低。此外，RIF-EV组孵化胚胎的侵袭能力显着降低。