PDF《两种方法冻存胎儿卵巢组织后的活力判断》

第29卷第3期2008年6月西安交通大学学报(医学版)Journal of Xiπan Jiaotong U niversity (Medical Sciences) Vol.

29 No.

3 J un.

2008 两种方法冻存胎儿卵巢组织后的活力判断 李跃萍1 ,邱曙东2 ,王燕蓉3 ,蔡玉芳3 ,沈新生3 ,崔岫3 ,黄元华1 (1. 海南医学院附属医院生殖医学中心 ,海南海口

570102 ;

2. 西安交通大学医学院人体解剖与组织胚胎学系 , 生殖医学研究中心 ,陕西西安

710061 ;

3. 宁夏医学院组织与胚胎学教研室 ,宁夏银川 750004) 摘要 :目的 比较常规慢速冻存法与快速冻存法保存胎儿卵巢组织的效果 ,为快速冻存法的应用提供依据.方法 以1.

5 mol/ L 乙二醇与 0.

1 mol/ L 蔗糖作冷冻保护剂 ,分别采用常规慢速冻存法和三步快速冻存法保存

1 mm3 大小 的胎儿卵巢组织 ,解冻后观察两组卵巢组织内的超微结构 ;

以去势裸鼠为受体 ,将解冻后的胎儿卵巢皮质片移植于裸 鼠的双肾被膜下 ,观察移植

2 周和

8 周后移植物的组织学结构、 移植物存活率、 卵泡密度及受体鼠血清 E2 水平. 结果 与新鲜胎儿卵巢组织相比 ,慢速和快速冷冻后卵巢组织中形态正常的卵母细胞的比例都明显减少 ( P <

0. 05) . 移植后

2 周的两移植组均有卵泡存活 ,原始卵泡广泛分布于基质中 ;

移植后

8 周的两移植组的卵泡数量都明显减少 , 但初级卵泡的数量相对增加.比较移植2 周和

8 周后移植物的存活率、 单位面积移植物内的原始卵泡或初级卵泡数 及受体鼠血清雌激素水平 ,两移植组并无显著性差异.结论 三步快速冻存法可有效保存胎儿卵巢组织的生物活 性 ,解冻后卵巢组织的形态学结构、 雌激素分泌与常规慢速冻存组并无显著性差异. 关键词 :胎儿 ;

卵巢 ;

深低温冻存 ;

移植 中图分类号 :R714 文献标识码 :A 文章编号 :167128259 (2008)

0320324205 Viability evaluation of human fetal ovarian tissue after frozen by two different freezing methods Li Yueping1 , Qiu Shudong2 , Wang Yanrong3 , Cai Yufang3 , Shen Xinsheng3 , Cui Xiu3 , Huang Yuanhua1 (1. Center for Reproductive Medicine , Affiliated Hospital of Hainan Medical College , Haikou

570102 ;

2. Research Center for Reproductive Medicine , Department of Histology and Embryology , Medical School of Xiπan Jiaotong University , Xiπan

710061 ;

3. Department of Histology and Embryology , Ningxia Medical College , Yinchuan

750004 , China) ABSTRACT : Objective To compare the effects of slow2rate f reezing method with that of rapid f reezing method f or cryop reservation of fetal ovarian tissue on viability of ovarian tissue and p rovide some data f or the application of rapid f reezing method. Methods Human fetal ovarian slices (1 mm3 in size) were f rozen with conventional slow cooling method and three2step rapid f reezing method , respectively , using f reezing solution containing 1.

5 mol/ L ethylene glycol and 0.

1 mol/ L sucrose. After thawed , electron microscopy was used to observe the ultrastructural changes in ovarian tissue. Ovariectomized female nude mice were chosen as recipients , two fetal ovarian tissue slices were separately inserted under kidney capsule of two sides in each mouse. The survival of ovarian grafts , histological structure , f ollicular density and serum E2 levels in recipients were detected after transplantation f or two or eight weeks. Results Compared with that of f resh ovarian tissue , the number of oocytes with normal morp hology in slow and rapid f reezing groups was significantly decreased ( P <