The Research Of Human Fetal Ovary Culture In Vitro

Objective: To evaluate whether the fresh and frozen-thawed human fetal ovary can develop and secret Estradiol (E₂) in tissue culture in vitro; to evaluate the influences of FSH and NAC on the featal ovary in vitro; to investigate the appropriate culture medium for human fetal ovary and offer some experimental data for the follwing study of human ovary transplantation.Methods: 30 human fetal ovaries of 20 to 28 weeks were cultured in vitro in 6 groups which culture medium had different factors. The culture medium was changed and stored for E₂ measure by radioimmunoassay (RIA) method every 3 days and the morphorlogic change of the cultured ovarian tissue was observed. Results: 1. Comparing with O and A groups, the E₂ output of the fetal ovary in B group was significantly greater, and the output increased with the longer culturing duration while decreased in O and A groups (P<0.05). 2. There was no significant difference of E₂ output and the proportion of more mature follcles between the fresh group and frozen-thawed group (P>0.05) while in both groups the output of E₂ and the proporation of more mature follicles significantly increased with the longer culturing duration (P<0.05). 3. Comparing with B group, the E₂ output and the proporation of more mature follicles of the fetal ovary in C, D and E groups were all significantly greater and the output and proporation of all increased with the longer culturing duration (P<0.05). 4.There was an obvious peak value of E₂ output every about 14—15 days which shows the periodity of E₂ secretion.Conclutions: 1. Human fetal ovary cultured in vitro could keep its histological structure and the ability of secreting E2, therefore, it may be used as the donor for ovary transplantation. 2. Human fetal ovary could endure the frozen-thawed process , therefore, it may broaden the source of donor for ovary transplantation and offer the necessary experimental data for the establishing of "Bank of ovary". 3. It was demonstrated that follicles in the cultured fetal ovary could develop further to more mature ones and this indicates that the secretion of E2 may mainly depends on the course of follicles development. 4. The culture medium containing FSH could enhance fetal ovary secreting E2 and improving the proporation of more mature follicles, which may depend on its ability of activating the hyperplasia of granulose cells and the synthesis of steroid hormone. 5. The culture medium containing NAC could enhance fetal ovary secreting E2 and improving the proporation of more mature follicles, which may depend on its ability of inhibiting the apoptosis of granulosa cells and atresia of follicles. 6. It was demonstrated that the culture medium containing FSH and NAC may act as a regular fetal ovary culture medium in vitro, since FSH and NAC could be in favor of keeping the activity of granulosa cells and increasing the E? output. 7. Human fetal ovary culture in vitro showed an obvious characteristic of fluctuation in E2 output, which may be related to the seasonal development and atresia of follicles in batches.