摘要
目的:探讨冷冻保存对人始基卵泡中颗粒细胞的形态及存活率的影响。方法:程序冷冻法保存人卵巢组织,采用常规组织学检测冷冻前后卵泡中颗粒细胞形态,并采用胶原酶消化联合镜下机械分离卵巢组织中的始基卵泡,采用双荧光标记,检测冷冻前后人卵巢组织分离始基卵泡中颗粒细胞的存活比例。结果:形态学检测发现,冷冻前卵巢组织中颗粒细胞发生损伤的始基卵泡占12.4%,卵母细胞损伤20.6%;冷冻后两者的比例分别为23.3%与24.4%。荧光标记的冷冻前卵巢组织中颗粒细胞存活的始基卵泡占72.7%,<50%颗粒细胞存活的卵泡仅占3.6%,卵母细胞存活率为85.5%;冷冻组中颗粒细胞存活的始基卵泡占79.5%,<50%颗粒细胞存活的卵泡仅占2.5%,卵母细胞存活率达86.1%。结论:冷冻复温的过程主要破坏卵泡中的颗粒细胞,而对卵母细胞的损伤很小。
Objective： To investigate the morphology and viability of granulosa cells in primordial follicles from fresh and cryopreserved human ovarian tissues. Methods： Two or three ovarian cortexes were randomly selected for histological examination from freshly obtained human ovarian tissue. The remaining ovarian material was incubated with collagenase type I and deoxyribonuclease I or performed to store in liquid nitrogen.After partial disaggregation,follicles were isolated by manual dissection. A double-staining method was used to assess granulose cells and oocyte viability. Results： In the fresh group, the viable percent of follicles with damaged granulosa cells and oocytes in tissue section were 12.4% and 20.6% compared with 23.3% and 24.4% in the frozen-thawed group by histological analysis.However,by double staining, 72.7%, 3.6% of freshly and 79.5%,2.5% of preservedly non-damaged granulosa cells and 〈50% of live granulosa cells in primordial follicles were viable by live-dead staining.When oocyte viability was considered alone, 85.5% and 86.1% of the cells were viable in fresh and frozen-thawed follicles, respectively. Conclusion：Granulosa cells, not oocytes of human primordial follicles are affected by the slowly-freezing procedure.
出处
《山东大学学报（医学版）》
CAS
北大核心
2005年第12期1112-1115,共4页
Journal of Shandong University:Health Sciences
基金
国家自然科学基金资助项目(39300136)
高等学校博士学科点专项科研基金资助课题(20030422033)
