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Problem statement: Development of efficient culture system to support embryonic development would be valuable when quality of produced embryos was important. However, the rate of bovine embryo production in vitro was still lower than expected. Present study, including of three experiments, was carried out to investigate the effect of insulin on nuclear maturation and subsequent development of immature bovine oocytes and in vitro fertilized embryos. Approach: Grade one cumulus-oocyte-complexes harvested from slaughterhouse ovaries were selected and randomly allocated in each treatment groups. In experiment 1, in vitro maturation medium (Hepes-buffered medium 199 + fetal calf serum + gonadotrophins + antibiotics) supplemented with 0 (control), 1, 10, 20 and 100 µg mL-1 of insulin. In experiment 2, to eliminate the effect of serum and hormones, Hepesbuffered medium 199 was supplemented with 1 mg mL-1 polyvinyl alcohols (PVA) and same levels of insulin. In experiment 3, the effect of insulin on bovine in vitro embryo development was assessed. Presumptive zygotes were randomly cultured in synthetic oviductal fluid added with 0 (control), 1, 10, 20 and 100 ìg mL-1 of insulin. Results: In experiment 1, nuclear maturation and embryo development rates were significantly higher in 1 and 10 µg mL-1 compared with other groups (P<0.05). In experiment 2, both maturation and cleavage rate significantly increased in 1 and 10 ìg mL-1 insulin. The only treatment resulted in higher hatchability was 10 ìg mL-1 insulin (17.1±2.34%) compared with control (11.34±3.94). In experiment 3, cleavage and morula rates were significantly greater in 1 and 10 µg mL-1 insulin compared with other groups; although the highest rates resulted by using 10 µg mL-1. Conclusion: Obtained results show that inclusion of 10 µg mL-1 insulin in maturation and culture medium exerted beneficial effects on nuclear maturation of bovine oocytes and in vitro embryo development till morula stage

Synergistic Effect of Insulin on <i>in vitro</i> Development of Immature Bovine Oocytes