Effects of supplementation of human endometriotic fluids on in vitro mouse preantral follicle culture

Abstract

This study aimed to investigate the potential detrimental effect of human endometriotic fluids (EFs) on in vitro growth of mouse preantral follicles. Preantral follicles (isolated from ovaries of 7- to 8-week-old mice) were cultured in growth medium for 10 days and then in maturation medium for 2 days. During in vitro culture, EF supernatants (0%, 2.5%, 5%, and 10%) were supplemented. Meiotic spindle integrity of metaphase II (MII) oocytes was analyzed. Hormone (17-estradiol and anti-Mullerian hormone [AMH]) levels in the final spent media were measured by enzyme-linked immunosorbent assay. The survival rates of follicles at day 10 were significantly lower in 3 EF-supplemented groups (56.1%, 30.6%, and 6.2%; 83.6% in the nonsupplemented group). The production of total oocytes per initiated follicle was also significantly lower in the 3 EF-supplemented groups (34.7%, 18.4%, and 4.1%; 68.1% in the nonsupplemented group). Proportions of the oocyte with normal spindles were significantly lower in the 3 EF-supplemented groups (10%, 0% and 0%; 52% in the nonsupplemented group). In the final spent media, the level of 17-estradiol was significantly lower only in the 10% EF-supplemented group, and the level of AMH was significantly lower in all 3 EF-supplemented groups, when compared with the nonsupplemented group. During in vitro culture of mouse preantral follicles, the survival rate, oocyte acquisition, spindle integrity of MII oocytes, and AMH production were greatly affected by EF supplementation. These findings suggest a possibility of detrimental effects of endometriotic cysts on folliculogenesis in adjacent ovarian tissues.