In vitro maturation of cynomolgus ovarian follicles = In vitro maturation of cynomolgus ovarian follicles

국문 초록 (Abstract)

목적: In vitro ovarian follicular growth (IVFG) is an important issue from the perspective of artificial ovary engineering and fertility preservation. Most of currently known IVFG conditions have been established using mice models, however, they have significantly different reproductive physiology compared to human. Therefore, it is necessary to use non-human primate models, which possess more similar characteristics of reproductive physiology to those of human. In this study, we tried to establish an optimal in vitro growth condition of cynomolgus monkey ovarian pre-antral follicles using different combination of gonadotropins.
방법: Under general anesthesia, ovaries of 3-year-old female cynomolgus monkey (Macaca fascicularis) were excised and collected. Pre-antral follicles were mechanically separated and the follicles containing layers of granulosa cells with a centrally located, visible oocyte, were selected for the experiments. These pre-antral follicles were cultured in droplets of 300 μL media containing nine types of FSH and LH combination and incubated for 14 days. Media was changed by refreshing half of the medium every other day. Morphological growth of the cultured follicles was evaluated under an inverted microscope and oocytes were matured further in vitro for 2 days in the presence of human chorionic gonadotropin (hCG).
결과: The characteristics of follicular growth were compared among the tested FSH and LH combination groups. In vitro grown follicles demonstrated visible expansion of granulosa and theca cells during culture period. The addition of 100-200 mIU FSH was superior to 10 mIU FSH. LH co-treatment showed better IVFG outcomes. From these groups, mature oocytes were successfully retrieved.
결론: Conclusively, we demonstrated cynomolgus monkey pre-antral follicles can be grown into mature follicles in vitro using specific FSH and LH combinations (HI14C2289).