大鼠卵巢组织冷冻复苏及自体异位移植研究  

A Study on the Heterotopic Autotransplantation Transplantation with Ovarian Tissues after Cryopreserved Thawed in Rats

作　　者：李冬秀[1] 耿杨柳[1] 吴小华 于晓惠[1] 张敏[1] 徐清华[1] 

机构地区：[1]解放军白求恩国际和平医院生殖医学中心,石家庄050082

出　　处：《解放军医药杂志》2015年第5期6-8,20,共4页Medical & Pharmaceutical Journal of Chinese People’s Liberation Army

基　　金：河北省计划生育委员会科研基金项目(2012-C02)

摘　　要：目的探讨直接覆盖玻璃化冷冻(direct cover vitrification,DCV)在冷冻大鼠卵巢组织中的效果,并对自体异位移植后卵巢组织存活状况及内分泌情况进行分析。方法性成熟的Wistar大鼠50只随机分为A、B、C、D组,A和B组各20只,C和D组各5只。A组为新鲜卵巢组织移植;B组卵巢组织经DCV冷冻保存2周复苏后移植;C组为去势对照;D组为假手术对照。自体异位移植6周后检测大鼠血清中雌二醇(E2)水平,计算卵泡密度,观察移植卵巢组织形态学及增殖细胞核抗原(proliferating cell nuclear antigen,PCNA)的表达情况。结果大鼠卵巢组织移植存活的卵巢组织周围可见血管供应,移植失败的卵巢组织呈纤维化状态,与周围组织血供不明显。B组卵巢组织移植存活率低于A组(P<0.05)。B组卵巢组织中卵泡密度均低于A、D组,且A组低于D组(P<0.05)。A、B组血清E2水平高于C组,低于D组(P<0.05)。A、B组卵巢组织PCNA阳性率均低于D组,且B组低于A组(P<0.05)。结论大鼠卵巢组织经DCV法冻融自体异位移植后部分卵泡能存活并可恢复内分泌功能。Objective To investigate the effect of direct cover vitrification（DCV） method in freezing ovarian tissues and to analyze the viability and endocrinic function after the autologous heterotopic transplantation in rats. Methods A total of 50 mature female rats were randomly divided into group A（ n = 20）,group B（ n = 20）,group C（ n =5） and group D（ n = 5）. Group A was transplanted with fresh tissues; group B was transplanted with cryopreserred thawed tissues after being cryopreserved for 2 weeks by DCV method; the bilateral ovarian tissues in group C were cut off,and group D underwent sham operations. After autologous heterotopic transplantation for 6 weeks,serum estradiol（ E2）levels were detected,and values of follicular density were calculated; histomorphology changes of transplanted ovaries and expressions of proliferating cell nuclear antigen（ PCNA） were observed. Results The vascularity was found in surrounding tissues of survival ovaries,while fibering and unconspicuous vascularity were found in unsuccessfully transplanted ovary tissues. The survival rate of grafts in group B was lower than that in group A（ P〈0. 05）. The value of follicular density in group B was lower than those in group A and D,and the value in group A was lower than that in group D（ P〈0. 05）. The serum E2 levels in group A and B were higher than that in group C,but the values were lower than that in group D（ P〈0. 05）. The positive rates of PCNA in group A and B were lower than that in group D,and the rate in group B was lower than that in group A（ P〈0. 05）. Conclusion Some ovarian tissues with autologous heterotopic transplantation after DCV and resuscitation method can survive and rehabilitate cover endocrine function.

关 键 词：卵巢 玻璃化作用 低温保存 移植物存活 移植 自体 大鼠 Wistar 

分 类 号：R715[医药卫生—妇产科学] R-332[医药卫生—临床医学]