Department of Biomedical and Molecular Sciences, Queen's University, Kingston, Ontario, K7L 3N6, Canada.
Department of Biomedical Sciences, Ontario Veterinary College, University of Guelph, Guelph, Ontario, N1G 2W1, Canada.
Sci Rep. 2017 Jan 12;7:40476. doi: 10.1038/srep40476.
Exosomes and microvesicles are extracellular vesicles released from cells and can contain lipids, miRNAs and proteins that affect cells at distant sites. Recently, microvesicles containing miRNA have been implicated in uterine microenvironment of pigs, a species with unique epitheliochorial (non-invasive) placentation. Here we report a novel role of conceptus-derived exosomes/microvesicles (hereafter referred to as extracellular vesicles; EVs) in embryo-endometrial cross-talk. We also demonstrate the stimulatory effects of EVs (PTr2-Exo) derived from porcine trophectoderm-cells on various biological processes including the proliferation of maternal endothelial cells (PAOEC), potentially promoting angiogenesis. Transmission immuno-electron microscopy confirmed the presence of EVs in tissue biopsies, PTr2-Exo and PAOEC-derived EVs (PAOEC-Exo). RT-PCR detected 14 select miRNAs in CD63 positive EVs in which miR-126-5P, miR-296-5P, miR-16, and miR-17-5P were the most abundant angiogenic miRNAs. Proteomic analysis revealed EV proteins that play a role in angiogenesis. In-vitro experiments, using two representative cell lines of maternal-fetal interface, demonstrated bidirectional EVs shuttling between PTr2 and PAOEC cells. Importantly, these studies support the idea that PTr2-Exo and PAOEC-Exo containing select miRNAs and proteins can be successfully delivered to recipient cells and that they may have a biological role in conceptus-endometrial cross-talk crucial for the pregnancy success.
外泌体和微泡是由细胞释放的细胞外囊泡，可包含影响远处细胞的脂质、miRNA 和蛋白质。最近，含有 miRNA 的微泡已被认为参与了猪的子宫微环境，猪具有独特的上皮绒毛膜（非侵入性）胎盘。在这里，我们报告了胚胎来源的外泌体/微泡（以下简称外囊泡；EVs）在胚胎-子宫内膜对话中的新作用。我们还证明了来自猪滋养层细胞的 EVs（PTr2-Exo）对包括母体内皮细胞（PAOEC）增殖在内的各种生物学过程的刺激作用，可能促进血管生成。透射免疫电子显微镜证实了组织活检、PTr2-Exo 和 PAOEC 衍生的 EVs（PAOEC-Exo）中存在 EVs。RT-PCR 在 CD63 阳性 EVs 中检测到 14 种选定的 miRNA，其中 miR-126-5P、miR-296-5P、miR-16 和 miR-17-5P 是最丰富的血管生成 miRNA。蛋白质组学分析揭示了在血管生成中起作用的 EV 蛋白。在体外实验中，使用母体-胎儿界面的两个代表性细胞系，证明了 PTr2 和 PAOEC 细胞之间双向 EVs 的穿梭。重要的是，这些研究支持这样的观点，即含有选定 miRNA 和蛋白质的 PTr2-Exo 和 PAOEC-Exo 可以成功递送到受体细胞，并且它们可能在胚胎-子宫内膜对话中发挥生物学作用，这对妊娠成功至关重要。
