玻璃化冷冻兔卵巢组织自体移植后卵巢功能观察

Observation of ovarian function after autologous implantation of vitrified ovarian tissues in female rabbits

目的观察玻璃化冷冻兔卵巢组织自体子宫系膜内种植后卵泡的发育情况。方法15只新西兰白兔随机分为两组:(1)冷冻复苏移植组,10只,切除双侧卵巢,将卵巢切成小组织块玻璃化冷冻2周后复苏,多点自体种植于子宫系膜内;(2)对照组,5只。通过卵巢组织光镜、阴道脱落细胞学检查,观察种植后卵巢组织长期存活、卵泡生长发育及内分泌功能恢复及维持情况;观察存活的卵巢组织对促性腺激素的反应性;采集MⅡ期成熟卵进行卵泡浆内单精子注射(intracytoplasmic sperm injection,ICSI),进一步观察移植后发育成熟的卵母细胞的受精情况。结果移植2个月和6个月后卵巢组织中各级卵泡所占的比例与正常卵巢组织相比差异无统计学意义(P>0.05);移植半年后对卵巢组织应用卵泡刺激素,与未用卵泡刺激素的卵巢组织相比,近成熟卵泡所占的比例增多(P<0.05);成熟卵母细胞ICSI受精后,能够得到正常的胚胎。结论玻璃化冷冻兔卵巢组织自体子宫系膜内种植能够很好地恢复卵巢功能,其功能可维持较长时间,结合体外受精技术可以产生正常的胚胎。

Objective To observe the follicle growth of vitrified rabbit ovarian tissues after autologous implantation in mesometrium in rabbits.Methods Fifteen female New Zealand rabbits were randomly divided into two groups.（1）Vitrified ovarian tissue group（n=10）： rabbits were bilaterally ovariectomized and the ovary was made into small tissues for vitrified freezing;two weeks later the frozen-thawed ovarian tissues were autologously transplanted into the mesometrium.（2）Control group included 5 animals.The long-term survival of ovarian tissues,survival and development of follicles,and endocrine function after gonadotropin treatment were evaluated by light microscope and vaginal exfoliocytology.The response of cryopreserved ovarian tissues to gonadotrophin was observed after autologous implantation.The oocytes of MⅡ stage were subjected to intracytoplasmic sperm injection（ICSI）to further observe the development potency of oocytes after autologous implantation.Results The ratios of follicles of each stage 2 months and 6 months after transplantation were similar to those of control group（P〈0.05）.Six months after follicle-stimulating hormone（FSH） stimulation,the ratio of mature follicles in ovarian tissues was significantly higher than that in the group untreated with FSH（P〈0.05）.The oocytes derived from ovarian tissues after superovulation could obtain normal embryos by ICSI.Conclusion Vitrified rabbit ovarian tissues can achieve ovarian function after autologous implantation in mesometrium,and the function can maintain for a long period;furthermore,normal embryos can be obtained by ICSI technique.