Kim Yong Bong, Ahn Sung Ho, Chang Doo Young, Chung Kyung Nam, Koh Jae Whoan
Department of Obstetrics and Gynecology, College of Medicine, InJe University, Seoul Paik Hospital, Seoul, Korea.
J Korean Med Sci. 2002 Apr;17(2):217-9. doi: 10.3346/jkms.2002.17.2.217.
Recent studies have suggested that the hydrosalpinx has a negative effect on pregnancy outcome, with markedly diminished implantation and increased early pregnancy loss. Fluid from the hydrosalpinx may leak into and accumulate in the uterine cavity. It is not clear, however if this creates a hostile local environment in the uterus for embryo implantation or exerts a direct embryotoxic effect. This study was conducted to investigate the detrimental effects of hydrosalpinx fluid (HSF) on the development of mouse embryos in vitro and to demonstrate whether Vero cells overcome these adverse effects. HSF was collected from three women with bilateral hydrosalpinx at the time of laparoscopic surgery. Collected fluid was centrifuged and the supernatant was frozen at -20 degrees C. For co-culture, Vero cells were commercially obtained in a frozen state and cultured using Ham's F10 medium. Single-cell mouse embryos (B6CBAF1) were cultured for 5 days in 0, 0.4, 0.8, and 1.2% of HSF in media with and without Vero cells and examined daily to record the number of embryos reaching expanded blastocyst and hatching stage. Co-culture of mouse embryos with Vero cells at 0.8% HSF concentration significantly enhanced embryo development, but not at 1.2% hydrosalpinx fluid concentration. These results suggest that HSF is highly embryotoxic and Vero cells are likely to overcome these detrimental effects to some degree.
近期研究表明,输卵管积水对妊娠结局有负面影响,会显著降低着床率并增加早期妊娠丢失率。输卵管积水的液体可能会漏入并积聚在子宫腔内。然而,尚不清楚这是否会在子宫内营造出不利于胚胎着床的局部环境,或者是否会产生直接的胚胎毒性作用。本研究旨在调查输卵管积水液(HSF)对小鼠胚胎体外发育的有害影响,并证明非洲绿猴肾细胞(Vero细胞)是否能克服这些不利影响。在腹腔镜手术时,从三名双侧输卵管积水的女性身上收集HSF。收集的液体进行离心,上清液在-20℃下冷冻。对于共培养,Vero细胞以冷冻状态购得,并使用哈姆氏F10培养基进行培养。单细胞小鼠胚胎(B6CBAF1)在含有和不含有Vero细胞的培养基中,分别在0%、0.4%、0.8%和1.2%的HSF中培养5天,并每天进行检查,记录达到扩张囊胚和孵化阶段的胚胎数量。在0.8% HSF浓度下,小鼠胚胎与Vero细胞共培养可显著促进胚胎发育,但在1.2%输卵管积水液浓度下则不然。这些结果表明,HSF具有高度胚胎毒性,而Vero细胞可能在一定程度上克服这些有害影响。