Adverse Effect of Human Hydrosalpingeal Fluid on the Developmentof Mouse Embryo

Koong, Mi Kyoung; Jun, Jin Hyun

Abstract

Recent studies reported that patients with hydrosalpinx have reduced pregnancy rates following in vitro fertilization and embryo transfer(IVF-ET). Hydrosalpingeal fluid(HSF) in the fallopian tube(s) can leak into the uterine cavity, and it may change the intrauterine microenvironment, and affect embryo development or implantation adversely. The objective of this study is to determine if HSF affects development or implantation adversely. The objective of this study is to determine if HSF affects development, hatching or implantation of mouse embryo in vitro. HSF was collected from 10 patients with hydrosalpinges undergoing salpingoneostomy. Collected fluid was centrifuged and supernatant was stored frozen at -20 C. For co-culture, human endometrial cells were obtained by endometrial biopsy and were cultured using Ham's F-10 medium. Two-cell mouse embryos(ICR-strain) were cultured for 6 days in T6 medium containing 0, 10, 20, 50 % of HSF with or without human endometrial cells. Statistical analysis was done by chi-square test. The ratios of developing mouse embryo in 0, 10, 20 50 % of HSF in media(n=80 in each group) were 70, 75, 70, 59* % to blastocyst, 70,65, 60, 46*% to hatching, and 50, 44, 48, 36 % to outgrowth, respectively. This mean ratios showed decreasing tendency according to increasing HSF concentration(*:p < 0.05). A similar tendency was observed in cluture with endometrial cell. The ratios of developing mouse embryo co-cultured with endometrial cells(n=40 in each group)were 82, 75, 65, 57* % to blastocyst, 78, 68, 63, 60 % to gatching, and 64, 55, 58, 58 % to implantation, respectively (*:p < 0.05). These data suggest that HSF adversely affect mouse embryo development and hatching rather than outgrowing or implantation in vitro. Thus, the hydrosalpinx-associated reduction in pregnancy rates among IVF-ET patients may be related, at least in part, to the deleterous effects of HSF on embryo development. In this study, the final medium concentrations were varying also by adding HSF, however, this dilution effect is considerd to be anlogous to in vivo condition when HSF influxes into the uterine fluid.