In Vitro Assessment of FSH and LH Effects on Progesterone Levels in Human Granulosa-Theca Cell Culture Running Title: Effect of FSH and LH on progesterone level in granulosa-theca cell culture

Abstract

Abstract The administration of LH just prior to the ovulation trigger could lead to new ideas for restore levels of excess progesterone level in the systemic circulation. This study aim to investigate the effect of applied r-LH on progesterone levels in human granulosa-theca tissue culture in response to various doses of administered r-FSH. The study enrolled 15 patients with normal menstrual cycles, aged between 18–38 in the reproductive period and underwent laparoscopic ovarian cystectomy at the Gynecology and Obstetrics department of Hacettepe University. Healthy ovarian tissue isolated from excised cyst tissue was cultured in vitro and basal levels (0th hour) of estrogen and progesterone release were determined. Afterwards, ovarian tissues were cultured with different concentrations of hormones in three groups, Group A;12.5 mIU/ml FSH; Group B;18.75 mIU/ml FSH; and Group C; 18.75 mIU/ml FSH + 6.25 mIU/ml LH. In the same way, hormone evaluation was performed at 24. and 48. hours, and the groups were compared. histopathological scoring was performed and immunofluorescence labeling was done using aromatase and cytochrome P450 for specific labeling of granulosa and theca cells, and Ki67 antibody was used to evaluate cell proliferation. Biochemical hormone analysis revealed no significant differences in the concentrations of progesterone or estrogen among the groups. Meanwhile, no significant differences were detected in the intensity measurements of aromatase, cytochrome P450 and Ki67 antibodies. Administering low or high dosages of FSH combined with LH therapy did not appear to have a significant impact on progesterone production.