Effects of follicular fluid-derived extracellular vesicles improve in vitro maturation and embryonic development of porcine oocytes

Heejae Kang, S. Bang, Heyyoung Kim, A. Han, Shuntaro Miura, Hye Sun Park, I. M. Saadeldin, Sanghoon Lee, Jongki Cho
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Abstract

To optimize the most efficient method for porcine in vitro maturation (IVM), we compared the effects of supplementing extracellular vesicles (EVs) derived from porcine follicular fluid (pFF). The cumulus oocyte complexes were grouped into 4 groups with different supplementations as following: pFF (G1), pFF-depleted EVs (G2), EVs (G3) and control (G4) groups. After IVM with different supplementations, maturation rates and the developmental competences of porcine oocytes and blastocyst development were investigated. Additionally, glutathione (GSH) and reactive oxygen species (ROS) levels were measured in mature oocytes. The EVs were isolated and characterized with cryo-TEM and nanoparticle tracking analysis. The pFF significantly affected the maturation rate, whereas the presence of EVs did not show notable difference in the maturation rates. Although there were numerical increases in the measured parameters in EV and pFF-depleted EVs groups, no significant differences were observed between them. The EV group showed similar oocyte maturation rate for both positive and negative control groups. The GSH was not different among the groups, but ROS levels were significantly lower in pFF-supplemented group when compared with other groups with the highest level in the control group. G2 group wasn’t significantly different G1 and G3 group. G3 group wasn’t significantly different from G2 and G4 group. This suggests that EVs in IVM medium which probably effected partially to protect against oxidative stress and potentially enhance the quality of oocytes. This study indicates that the EVs in pFF play a significant role in improving the efficiency of oocyte maturation in porcine.
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卵泡液衍生的细胞外囊泡对改善猪卵母细胞体外成熟和胚胎发育的影响
为了优化猪体外成熟(IVM)的最有效方法,我们比较了补充猪卵泡液(pFF)提取的细胞外囊泡(EVs)的效果。将积液卵母细胞复合体分为 4 组,分别添加不同的补充物:pFF 组(G1)、pFF-depleted EVs 组(G2)、EVs 组(G3)和对照组(G4)。用不同的补充剂进行 IVM 后,研究了猪卵母细胞的成熟率和发育能力以及囊胚的发育情况。此外,还测量了成熟卵母细胞中谷胱甘肽(GSH)和活性氧(ROS)的水平。通过冷冻-TEM和纳米粒子跟踪分析分离并鉴定了EVs。pFF 明显影响成熟率,而 EVs 的存在对成熟率没有明显影响。虽然 EV 组和 pFF 贫化 EV 组的测量参数在数值上有所增加,但它们之间没有观察到明显的差异。在阳性对照组和阴性对照组中,EV 组的卵母细胞成熟率相似。各组之间的 GSH 没有差异,但补充 pFF 组的 ROS 水平与其他组相比明显较低,其中对照组的 ROS 水平最高。G2 组与 G1 和 G3 组无明显差异。G3 组与 G2 和 G4 组无明显差异。这表明,IVM 培养基中的 EVs 可能对氧化应激有部分保护作用,并有可能提高卵母细胞的质量。这项研究表明,pFF 中的 EVs 在提高猪卵母细胞成熟效率方面发挥了重要作用。
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来源期刊
Korean Journal of Veterinary Research
Korean Journal of Veterinary Research Veterinary-Veterinary (all)
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