Responce Of Cryopreserved Ovarian Tissue After Autologous Implantation In Mouse Stimulated With Gonadotrophin

OBJECTIVE: To investigate the response of cryopreserved ovarian tissue after autologous implantation in mouse stimulated with gonadotrophin.METHODS: Thirty six female mice were randomly divided into three groups, with 12 mice in each group. In group of fresh ovarian tissue , each ovary was cut into there pieces , the volum of each piece was about 1 mm×2 mm×2 mm, fresh ovarian tissue was implanted into kidney capsule of mice; in group of cryopreserved ovarian tissue, each ovary was cut into there pieces ,ovarian tissue was implanted into kidney capsule of mice after cryopreserved by vitrification for two weeks. We investigate the response of ovarian tissue after autologous implantation stimulated with gonddotrophin. Two weeks later after autologous implantation , six mouse were picked out from each group , to investigate the response of ovarian tissue stimulated with gonadotrophin, 7. 5IU Human menopausal gonadotropin and 10IU Human chorionic gonadotropin were used . The ovarian histology, the survival and development of follicles and the response of cryopreserved ovarian tissue after autologous implantation in mouse stimulated with gonadotrophin were detected through light microscope and counting the follicles of every stages. Vaginal cytology was used to assess the survival of implanted ovarian tissues. Immunohistochemistry staining method was used to observe the expression of follicle stimulating hormone receptor. We picked twenty preantral follicles and twenty antral follicles from each group, to gather pictures under the same condition. The Image-Pro Plus5. 0 software wes used to obtain the integrated opticaldensity of follicle stimulating hormone receptor expression.RESULTS: Before and after stimulation with gonadotrophin, themature follicle rate of group of fresh ovarian tissue was 2.3% and 4. 2%, that of group of cryopreserved ovarian tissue was 2. 3% and 4. 0%, and that of group of control was 2. 6% and 5. 8%. Regarding the percentages of mature follicle, there were significant statistically after stimulated with gonadotrophin ( P<0. 05) ; After stimulating with gonadotrophin the percentages of mature follicle were the same in the fresh tissue group, cryopreserved tissue group and control group ( P>0. 05) ; The integrated optical density of follicle stimulating hormone receptor of fresh ovarian tissue were 9408 + 2777 and 4531±1903 of cryopreserved ovarian tissue were 9174 + 3092 and 4808 + 1385, and of the control ovarian tissue were 8838 + 2064 and 5516+1136 respectively. The integrated optical density of follicle stimulating hormone receptor expression of the there groups compared one by one. There was no significant difference between any two groups ( P>0. 05) .CONCLUSIONS: The follicle stimulating hormone receptor is preserved by cryopreservation and transplantation, after cryopreservation and ectopic transplantation small pieces of ovarian tissue response to gonadotropin stimulation is normal.