Kim Minji, Park Ji-Eun, Lee Yongjin, Lee Seung-Tae, Lee Geun-Shik, Hyun Sang-Hwan, Lee Eunsong, Lee Joohyeong
Laboratory of Theriogenology, College of Veterinary Medicine, Kangwon National University, Chuncheon 24341, Gangwon, Republic of Korea.
Division of Applied Animal Science, College of Animal Life Science, Kangwon National University, Chuncheon 24341, Gangwon, Republic of Korea.
Animals (Basel). 2023 Mar 30;13(7):1206. doi: 10.3390/ani13071206.
This study evaluated the effect of various growth factors and hormones in an in vitro growth (IVG) medium on the in vitro maturation (IVM) and developmental competence of oocytes derived from small antral follicles (SAFs) in pigs. Cumulus-oocyte complexes (COCs) derived from SAFs were either untreated or treated with epidermal growth factor (EGF), insulin-like factor-1 (IGF-1), insulin, or growth hormone (GH) for 2 days of IVG. Following IVG, COCs were cultured for maturation, and IVM oocytes were induced for parthenogenesis (PA). During IVG, the nuclear maturation of oocytes was significantly increased by the insulin treatment compared to other treatments. Moreover, the insulin treatment significantly increased blastocyst formation after PA relative to the No-IVG, control, EGF, and GH treatments. The cumulus expansion score after IVG-IVM was significantly higher in the insulin group than in the other groups. The glutathione (GSH) contents in IVM oocytes were increased through treatment with IGF, insulin, and GH compared to those of No-IVG oocytes. The level of reactive oxygen species (ROS) in IVM oocytes in all treatment groups was significantly lower after IVG culture than in the No-IVG group. The maturation-promoting factor (MPF) activity after IVM in the insulin-treated oocytes was significantly higher than that of the oocytes treated with EGF, IGF-1, and GH. In conclusion, this study demonstrates that insulin treatment during IVG culture improves the maturational and developmental competence of oocytes derived from SAFs in pigs through its effect on cumulus cell expansion and cytoplasmic microenvironments, such as GSH, ROS, and MPF activity.
本研究评估了体外生长(IVG)培养基中各种生长因子和激素对猪小窦卵泡(SAF)来源卵母细胞的体外成熟(IVM)和发育能力的影响。从SAF获得的卵丘-卵母细胞复合体(COC)在IVG培养2天时,要么不进行处理,要么用表皮生长因子(EGF)、胰岛素样因子-1(IGF-1)、胰岛素或生长激素(GH)进行处理。IVG后,将COC培养用于成熟,并诱导IVM卵母细胞进行孤雌生殖(PA)。在IVG期间,与其他处理相比,胰岛素处理显著提高了卵母细胞的核成熟率。此外,相对于未进行IVG、对照、EGF和GH处理,胰岛素处理显著增加了PA后的囊胚形成率。IVG-IVM后的卵丘扩展评分在胰岛素组显著高于其他组。与未进行IVG的卵母细胞相比,通过IGF、胰岛素和GH处理,IVM卵母细胞中的谷胱甘肽(GSH)含量增加。IVG培养后,所有处理组IVM卵母细胞中的活性氧(ROS)水平均显著低于未进行IVG组。胰岛素处理的卵母细胞在IVM后的成熟促进因子(MPF)活性显著高于用EGF、IGF-1和GH处理的卵母细胞。总之,本研究表明,IVG培养期间的胰岛素处理通过其对卵丘细胞扩展和细胞质微环境(如GSH、ROS和MPF活性)的影响,提高了猪SAF来源卵母细胞的成熟和发育能力。