Effect of Growth Factors and Hormones during In Vitro Growth Culture of Cumulus-Oocyte-Complexes Derived from Small Antral Follicles in Pigs

SIMPLE SUMMARY: This study investigated the effect of various growth factors and hormones in an in vitro growth medium on in vitro maturation and the developmental competence of oocytes derived from small antral follicles in pigs. Cumulus–oocyte complexes derived from small antral follicles (<3 mm) were cultured in an in vitro growth medium supplemented with epidermal growth factor (EGF), insulin-like factor-1 (IGF-1), insulin, or growth hormone (GH). The effects on oocyte growth, nuclear progression, oocyte maturation, and embryonic development after parthenogenesis were examined. Insulin treatment during oocyte growth was found to have a positive effect on nuclear maturation, embryonic development after parthenogenesis, and the expansion of cumulus cells. Insulin showed positive results by enhancing cytoplasmic maturation, decreasing free radical content, and increasing maturation-promoting factor activity. ABSTRACT: 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.