P-23: Effect of Melatonin on In vitro Maturation, Fertilization and Resulted Embryo Development in Mouse

Background: Melatonin has been recognised as effective reactive oxygen species (ROS) scavenger. antioxident melatonin effects has been expressed in management of cancer, alzheimer, diabet diseases. This study designed to determine the effect of melatonin on in vitro oocytes maturation and fertilization and cleavage and blastocyst development rate of mouse. Materials and Methods: Cumulus - oocyte complex (COCs) and germinal vesicle (GV) were obtained from 6-8 weeks female NMRI mice 46-48 hours after administration of an intra-peritoneal injection of 5 IU of PMSG. COCs were cultured for 18 hours in maturation medium (TCM199), supplemented with different doses of melatonin (100, 10, 1μM, 100, 10μM) and control group (without melatonin) and their maturation rate was assessed. To assay melatonin effect on embryo development rate, after in vitro fertilization, two pronucleous embryos were culture in T6 medium supplemented with different doses of melatonin and control group (without melatonin) and embryos development rate was recorded until blastocyst stage by inverted microscope. Also, two-cell embryos were obtained from oviduct 48 hours after administration of an intra-peritoneal injection of 5 IU pregnant mares’ serum gonadotrophin and subsequent human chorionic gonadotrophin (ip). Embryos were cultured in T6 supplemented with different dosages of melatonin {0 (control group), 100 µM/ml, 10 µM/ml, 1 µM/ml, 100 nM/ml and 10nM/ml (1-5 treatment groups) . In notice to embryos percent in different stages, the rate of embryos development was assessed using of invert microscope and compared to control group. Outcomes were analyzed using Chi-square test for SPSS version 13.0. Results: Our data indicate that 1, 10 and 100μM of melatonin has a positive effect on mouse in vitro oocyte maturation and fertilization in comparison to control group (p<0.01). Also, the cleavage and blastocysts development rate increased in presence of 10and 100 nM of melatonin (p<0.01). The results showed that the rate of cleavage and development of mouse embryos increased significantly in the development culture medium supplemented with 10 and 100 nM/mg of melatonin in comparison to control group (p<0.001). Conclusion: Melatonin as effective ROS scavenger has positive influence on in vitro oocytes maturation and embryos development promotion of mouse.