Pérez-García Felipe, Muñoz-Acuña Erwin, Valencia Cecilia, Aguila Luis, Felmer Ricardo, Arias María Elena
Laboratory of Reproduction, Centre of Reproductive Biotechnology (CEBIOR-BIOREN), Faculty of Medicine, Universidad de La Frontera, Temuco 4811230, Chile.
Doctoral Program in Sciences, Major in Applied Cellular and Molecular Biology, Faculty of Agriculture and Environmental Sciences, Universidad de La Frontera, Temuco 4811230, Chile.
Int J Mol Sci. 2025 Mar 22;26(7):2880. doi: 10.3390/ijms26072880.
Small extracellular vesicles (sEVs) play a crucial role in intercellular communication and have demonstrated significant relevance in reproductive biotechnology, particularly in in vitro maturation (IVM) and bovine embryo production. This study evaluates the effects of bovine follicular fluid-derived extracellular vesicles (ffsEVs) isolated using two methods: ultracentrifugation (UC) and size-exclusion chromatography (SEC) on oocyte maturation and preimplantational embryonic development. Significant differences in the size of ffsEVs obtained by both isolation methods were noted, with UC-derived ffsEVs (UC ffsEVs) being smaller than those isolated by SEC (SEC ffsEVs). UC ffsEVs were more effective in upregulating critical oocyte quality genes, such as and . However, no significant differences were observed in embryonic developmental rates. Furthermore, the expression of genes associated with preimplantational embryonic quality revealed that only the SEC ffsEVs group exhibited a significant increase in and levels, indicating an enhancement in embryonic quality. Notably, blastocysts derived from SEC ffsEVs also showed a higher total cell count compared to those from UC ffsEVs. No differences were found in other critical genes like and . These results suggest that the use of SEC ffsEVs could improve the in vitro embryo production process, highlighting the importance of the isolation method in determining the functional efficacy of ffsEVs according to research objectives.
小细胞外囊泡（sEVs）在细胞间通讯中发挥着关键作用，并且在生殖生物技术中已显示出显著相关性，特别是在体外成熟（IVM）和牛胚胎生产方面。本研究评估了使用两种方法分离的牛卵泡液衍生的细胞外囊泡（ffsEVs）对卵母细胞成熟和植入前胚胎发育的影响，这两种方法分别是超速离心（UC）和尺寸排阻色谱法（SEC）。两种分离方法获得的ffsEVs大小存在显著差异，超速离心法获得的ffsEVs（UC ffsEVs）比尺寸排阻色谱法分离的（SEC ffsEVs）更小。UC ffsEVs在上调关键卵母细胞质量基因（如 和 ）方面更有效。然而，在胚胎发育率方面未观察到显著差异。此外，与植入前胚胎质量相关的基因表达表明，只有SEC ffsEVs组的 和 水平显著增加，表明胚胎质量有所提高。值得注意的是，与UC ffsEVs相比，SEC ffsEVs来源的囊胚总细胞数也更高。在其他关键基因（如 和 ）方面未发现差异。这些结果表明，使用SEC ffsEVs可以改善体外胚胎生产过程，突出了根据研究目标确定ffsEVs功能功效时分离方法的重要性。
