The potentiality of two-dimensional preantral follicle culture as an in vitro model in predicting premature ovarian failure

The purpose of this study is to identify the potential of a two-dimensional preantral follicle culture as an in vitro model of predicting premature ovarian failure . The two-dimensional preantral follicles culture method was established by cultivating preantral follicles collected from ICR F1 hybrids (aged 12–14   days) for 12   days. The preantral follicles were incubated with 0.54   mg/ml cyclophosphamide , 0.5   mg/ml busulfan , 0.12   mg/ml cisplatin , 3.12   mg/ml 4-vinylcyclohexene diepoxide, 5   mg/ml D (+) galactose , and 0.5   mg/ml hydrocortisone for 24   h at culture days 2, 6 and 11. The diameter of follicles, the cumulus-oocyte complex number and the maturity of oocytes were recorded as the parameters to detect follicular maturation induced by the culture agents. The results indicated that, except for busulfan, D (+) galactose, and hydrocortisone, such test articles could significantly decrease follicular growth ( p   <   0.05 or p   <   0.01), and induce oocyte degeneration and follicle atresia when the follicles were treated at day 2. With the exception of hydrocortisone, such agents also gradually decreased follicular development ( p   <   0.05 or p   <   0.01) when the follicles were treated at day 6. All of the test articles but hydrocortisone can interfere with the ovulation , the cumulus-oocyte complex discharge and oocyte maturation of follicles when treated at days 2, 6 and 11. It is suggested that two-dimensional preantral follicle culture could be utilized as a potential in vitro system to mimic the POF model. It may also be employed in screening potential ovarian toxic agents, reducing laboratory animal use and promoting animal welfare .