Department of Theriogenology and Biotechnology, College of Veterinary Medicine, Seoul National University, Seoul 08826, Republic of Korea.
Department of Microbiology and Immunology, School of Medicine, Eulji University, Daejeon 35233, Republic of Korea.
Int J Mol Sci. 2023 Jun 6;24(12):9807. doi: 10.3390/ijms24129807.
Several studies have examined exosomes derived from porcine follicular fluid (FF), but few have reported their application in controlled experiments. The main concern in the field of embryology may be that controlled conditions, such as using a defined medium intermittently, cause poor results in mammalian oocyte maturation and embryo development. The first reason is the absence of the FF, which copes with the majority of the processes emerging in oocytes and embryos. Therefore, we added exosomes derived from porcine FF to the maturation medium of porcine oocytes. For morphological assessment, cumulus cell expansion and subsequent embryonic development were evaluated. Moreover, several stainings, such as glutathione (GSH) and reactive oxygen species (ROS), fatty acid, ATP, and mitochondrial activity, as well as evaluations of gene expression and protein analysis, were used for the functional verification of exosomes. When the oocytes were treated with exosomes, the lipid metabolism and cell survival of the oocytes were fully recovered, as well as morphological evaluations compared to the porcine FF-excluded defined medium. Therefore, controlled experiments may produce reliable data if the exosomes are treated with the desired amounts, and we suggest applying FF-derived exosomes to promote experimental data when performing controlled experiments in embryology.
已有多项研究对猪卵泡液(FF)衍生的外泌体进行了检测,但很少有研究报道其在对照实验中的应用。胚胎学领域的主要关注点可能是,在哺乳动物卵母细胞成熟和胚胎发育过程中,使用定义明确的培养基间歇性处理等对照条件会导致结果不佳。第一个原因是缺乏 FF,它应对卵母细胞和胚胎中出现的大多数过程。因此,我们将猪 FF 衍生的外泌体添加到猪卵母细胞的成熟培养基中。为了进行形态评估,评估了卵丘细胞的扩张和随后的胚胎发育。此外,还进行了几种染色,如谷胱甘肽(GSH)和活性氧(ROS)、脂肪酸、ATP 和线粒体活性,以及基因表达和蛋白质分析评估,以对外泌体的功能进行验证。当用外泌体处理卵母细胞时,与排除猪 FF 的定义明确培养基相比,卵母细胞的脂质代谢和细胞存活得到了完全恢复,并且进行了形态评估。因此,如果以所需量处理外泌体,对照实验可能会产生可靠的数据,我们建议在胚胎学中进行对照实验时应用 FF 衍生的外泌体来促进实验数据。