冻融后移植的小鼠卵巢组织对促性腺激素的反应

Response of cryopreserved ovarian tissue after autologous implantation in mouse stimulated with gonadotrophin

目的探讨冻融后移植的小鼠卵巢组织对促性腺激素的反应。方法将36只性成熟雌性小白鼠随机分为新鲜移植组、冻融移植组和对照组，每组12只。新鲜移植组小鼠切除双侧卵巢，将卵巢切成小组织块，立即移植入双侧肾被膜下；冻融移植组小鼠切除双侧卵巢，将卵巢切成小组织块，采用玻璃化冷冻方法冷冻保存，2周后将冷冻卵巢组织复苏，移植入小鼠双侧肾被膜下。卵巢组织移植2周后，新鲜移植组和冻融移植组每组随机取6只小鼠应用7．5IU人绝经期促性腺激素及10IU绒毛膜促性腺激素，观察移植后的卵巢组织对促性腺激素的反应。同时应用免疫组化染色方法观察各组卵泡中卵泡刺激素受体的表达情况。结果新鲜移植组、冻融移植组和对照组未应用促性腺激素小鼠卵巢组织内近成熟卵泡百分率分别为2．3％、2．3％和2．6％，应用促性腺激素小鼠卵巢组织内近成熟卵泡百分率分别为4．2％、4．0％和5．8％，各组内分别比较，差异均有统计学意义（P〈0．05）；新鲜移植组和冻融移植组与对照组比较，差异均无统计学意义（P〉0．05）。新鲜移植组、冻融移植组和对照组卵泡刺激素受体表达积分吸光度值在窦状卵泡中分别为9408±2777、9175±3093和8838±2064，在窦前卵泡中分别为4531±1903、4808±1386和5516±1136，各组间分别比较，差异均无统计学意义（P〉0．05）。结论卵巢组织冷冻保存、复苏及移植过程未影响卵巢卵泡刺激素受体的表达，冻融后移植的小鼠卵巢组织对外源性促性腺激素的反应未受冷冻、复苏及移植等过程影响。

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, fresh ovarian tissue was implanted into kidney capsule of mice; in group of cryopreserved ovarian tissue, ovarian tissue was implanted into kidney capsule of mice after cryopreserved by vitrification for two weeks. We investigated the response of ovarian tissue two weeks later after autologous implantation stimulated with gonadotrophin. Immunohistochemistry staining method was used to observe the expression of follicle stimulating hormone receptor. Results Before and after stimulation with gonadotrophin, the mature 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 differences after stimulation 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 in antrofollicle and pre-antrofollicle were 9408 ±2777 and 4531 ± 1903, that of cryopreserved ovarian tissue were 9175 ± 3093 and 4808 ± 1386, and that of the control ovarian tissue were 8838 ±2064 and 5516 ± 1136 respectively. There was no significant difference between any two groups （P 〉 0.05）. Conclusion The follicle stimulating hormone receptor is preserved by cryopreservation and transplantation, small pieces of ovarian tissue response to gonadotropin stimulation is normal.