Development of Mouse Preantral Follicle after In Vitro Culture in A Medium Containing Melatonin

OBJECTIVE Improvements in cancer treatment have allowed more young women to survive. However, many cancer patients suffer from ovarian failure. Cryopreservation is one of the solutions for fertility restoration in these patients. The cryopreservation of isolated follicles is a more attractive approach in the long term. Many endocrine and paracrine factors can stimulate the granulosa cells of preantral follicles to proliferate. Melatonin acts as direct free radical scavenger and indirect antioxidant. In this study, we investigated the direct effects of melatonin on follicle development and oocyte maturation by exposing in vitro cultured mouse vitrified-warmed ovarian follicles to melatonin. MATERIALS AND METHODS In an experimental study, preantral follicles with diameter of 150-180 µm were isolated from prepubertal mouse ovaries. Follicles were vitrified and thawed using cryolock method. They were then cultured individually for 7 days in droplets supplemented with 0, 10 and 100 pM melatonin, while ovulation was induced using epidermal growth factor (EGF) and human chorionic gonadotropin (hCG). The survival rate of follicles and nuclear maturation of ovulated oocytes were determined. RESULTS At the end of culture, significant increases in follicle survival (p<0.001) and in diameter (p<0.05) were noticed in 10 pM melatonin group compared to control group. In the 100 pM group, survival rate was not affected by melatonin. It was revealed that after induction of ovulation, total number of metaphase II oocytes in treatment groups were not influenced by melatonin (p>0.05). CONCLUSION Culture of mouse vitrified-warmed preantral follicles in a medium supplemented with 10 pM melatonin increased the number of surviving follicles.