Extracellular vesicles from seminal plasma to improve fertilizing capacity of bulls.

Anna Lange-Consiglio, Emanuele Capra, Noemi Monferrini, Simone Canesi, Giampaolo Bosi, Marina Cretich, Roberto Frigerio, Valentina Galbiati, Federica Bertuzzo, Francesco Cobalchini, Fausto Cremonesi, Bianca Gasparrini
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Abstract

Seminal plasma contains extracellular vesicles (EVs) that vehicle RNA, proteins, and other molecules able to influence the biological function of sperm. The aim of this study was to improve the fertilizing capacity of male gametes of low-fertility bulls using EVs isolated by ultracentrifugation from the seminal plasma of a bull of proven fertility. After dose-response curve, 10×106 sperm of low-fertility bulls were co-incubated for an hour with 400×106 EVs/ml. In addition, it has been verified that the incorporation of EVs, which takes place in the sperm midpiece, is maintained for 5 hours and even after cryopreservation. Subsequently, the spermatozoa of low-fertility bulls, with EVs incorporated, were used for the in vitro production of embryos. The rate of blastocyst at seventh day yield in vitro, with the use of sperm with EVs incorporated, increased by about twice the yield obtained with the same sperm in the absence of EVs: bulls having an average embryonic yield of 6.41±1.48%, 10.32±4.34% and 10.92±0.95% improved their yield to 21.21±1.99%, 22.17±6.09% and 19.99±5.78%, respectively (P<0.05). These encouraging results suggest that it might be possible to keep breeding bulls with poor fertility. Further studies will be needed to evaluate the in vivo fertility of sperm treated with EVs and understand how the content of EVs is involve in the sperm-vesicle interaction and in the improved sperm performance.

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从精浆中提取细胞外囊泡,提高公牛的受精能力。
精浆中含有细胞外囊泡 (EV),这些囊泡中含有 RNA、蛋白质和其他能够影响精子生物功能的分子。本研究的目的是利用从经证实具有生育能力的公牛精浆中通过超速离心分离出来的 EVs 提高低生育力公牛雄性配子的受精能力。在剂量反应曲线上,10×106 个低繁殖力公牛的精子与 400×106 个 EVs/ml 共同孵育一小时,结果表明,EVs/ml 对低繁殖力公牛的受精能力有显著提高。此外,经证实,EVs 在精子中段的结合可维持 5 小时,甚至在冷冻保存后也是如此。随后,加入了 EVs 的低繁殖力公牛精子被用于体外生产胚胎。使用含有 EVs 的精子后,体外第七天的囊胚率比未含有 EVs 的精子提高了约两倍:公牛的平均胚胎率分别为 6.41±1.48%、10.32±4.34% 和 10.92±0.95%,胚胎率分别提高到 21.21±1.99%、22.17±6.09% 和 19.99±5.78%(P<0.05)。
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