冻融人胎儿卵巢组织异种移植后早期血管生成的实验观察  

Experimental observation on angiogenesis of the frozen-thawed human fetal ovaries after xenotransplantation

作　　者：吴丹丹[1] 熊成容[1] 林海燕[1] 徐炳森[1] 童月红[1] 叶碧绿[1] 林金菊[1] 周颖[1] 

机构地区：[1]温州医学院附属第一医院生殖医学中心

出　　处：《解剖学报》2009年第1期113-117,158,共6页Acta Anatomica Sinica

基　　金：国家自然科学基金项目(30370749);浙江省科技厅项目(2003C33001)资助

摘　　要：目的观察冻融人胎儿卵巢组织异种移植后1个月内移植物形态学改变，动态了解微血管情况。方法将难免流产胎儿的卵巢组织经过冻融，异种移植至18只裸鼠肾被膜下，分别于移植后2d、7d和28d回收移植物。将新鲜、冻融以及移植后不同时间的卵巢组织进行光镜、电镜观察，RT-PCR检测血管相关基因的表达。结果移植后2d卵巢组织的微血管密度显著上升，部分血管处于扩张状态；移植后7d微血管密度达到相对高峰且扩张的血管腔消失，微小血管恢复正常形态；移植后28d微血管密度出现轻微下降。电镜观察发现，移植卵巢内多数原始卵泡结构完整；移植后2d间质水肿，血管内红细胞淤积；移植后7d及28d的问质水肿逐渐消退，大量微小血管生长。移植卵巢的人血管内皮生长因子（VEGF）和血管生成素2（Ang-2）mRNA表达均呈双向改变，即在移植后2d上升到高值，此后表达下降。结论冻融人胎儿卵巢组织移植后的血管重建在移植后1个月内基本完成，移植后1周是采取措施提高冻融人胎儿卵巢移植效果的关键时期。Objective To observe the angiogenesis in the human fetal ovarian grafts one month after heterotrausplantation. Methods Ovaries from 3 inevitable abortion human fetuses 22 gestational weeks of age were frozen- thawed, xenografted into the immunodeficient nu/nu mice （n = 18 ）, and recovered on 2, 7, and 28 days after transplantation respectively. These grafts, together with the fresh and the frozen-thawed ovarian tissue were examined for blood vessels by immunolocalization of CD34, ultrastructure, and detection of the mRNA expression of vascular endothelial growth factor （VEGF）, Angiopientin-2（Ang-2）. The untransplanted tissues of the fresh and the frozen-thawed ovarian were investigated exactly the same as the grafts. Results The density of the microvessel was significantly increased in the grafts on day 2, peaked on day 7, and slightly reduced on day 28, together with distended microlvessel on day 2 which turned to normal on day 7 under the light Microscope. Transmission electron microscope （TEM） showed the majority primordial follicles were well preserved. The grafts of day 2 showed some edema in stroma and vessels filled with red blood cells. As they disappeared, the microvessel was growing in the counterparts on day 7 and day 28. The mRNA levels of VEGF and Aug-2 showed the similar temporary rising on day 2 and decreasing since. Conclusion The angiogenesis of the frozen-thawed ovarian tissue from human fetus completed within one month after transplantation. The first week after transplantation was the critical time to achieve prolonged graft functionality and increased the yield of oocytes.

关 键 词：卵巢 冻融 异种移植 血管生成 反转录-聚合酶链反应 胎儿 人 

分 类 号：R339.2[医药卫生—人体生理学]