人卵巢组织冷冻保存的实验研究

目的 :探讨冷冻保存对人卵巢皮质内各级卵泡形态及对组织体外培养的影响。方法 :取手术切除的 30例患者的卵巢皮质 ,放入二甲基亚砜配制的冷冻保护液中 ,应用程序冷冻法保存于液氮内。冷冻前后取部分卵巢皮质行组织学检测 ;同时培养部分组织块观察上清中雌二醇分泌的变化。结果 :冷冻后存活的始基卵泡 ,与新鲜组比较无显著改变 (P >0 .0 5 ) ,初级卵泡的存活率显著下降 (P <0 .0 5 ) ;而冷冻保存不会明显降低始基卵泡与初级卵泡总的存活率 (P >0 .0 5 )。复温后卵巢组织中仅见 1枚形态正常的次级卵泡。冷冻后培养上清中雌二醇水平与新鲜组比较无明显差异 (P >0 .0 5 )。结论 :程序冷冻法可以较好地保存人卵巢皮质内的始基卵泡 ,且不影响冷冻后卵巢组织的激素分泌。

The morphological study of cryopreserved human ovarian cortical tissue

Objective:To investigate the effects of frozen-th awed process on the m orphology of various stage follicles and human ovarian cortical tissue in vitro culture.Methods:Pieces of human ovarian cortex were frozen slo wly in program mable freezer and then plunged into the liquid nitrogen. The cryopreserved ovari an tissues were thawed in 37℃ water bath and washed free of dimethyl sulfoxid e.Two or three fragments were randomly selected and fixed for morphologic al exa mination from both fresh and stored ovarian tissues.Follicular development and estradial secretion in cultured ovarian cortexes were calculated and compared wi th those in non-cultured tissue.Results:Morphological examin ation indicated t hat 75.19%(200/266),21.80%(58/266),3.01%(8/266) of primordial,primary and sec ondary follicles,respectively,were in slices of fresh tissue,these proportion s were altered by 79.31%(207/261),19.16%(50/261),1.53%(4/261) after cryopreser vation.In the fresh group,the percent viability of primordial follicle and pri mary follicle were 83.49%±21.01% and 77.84%±19.57%,respectively,compared wit h 7 6.14%±28.31%(P>0.05)and 61.06%±25.41%(P <0.05)。However,the number of both healthy primordial and healthy primary follicle in fresh ovarian tissue was not different from that in frozen group(P>0.05).Only one healthy secondary follic le was found in the cryopreserved ovarian cortical tissue.Levels of E 2 secrete d in fresh ovarian cortical fragments were not significantly different from those in frozen ovarian tissues.Conclusion:Cryopreserved human ova rian tissue can m antain similar primordial follicle viability and E 2 secretion with fresh ovari an cortex.