冻融人胎儿卵巢组织异种移植的实验研究  

The experimental study on the xenograft of frozen-thawed human fetal ovarian tissue

作　　者：韩海艳[1] 周颖[2] 金畅[3] 傅晓艳[1] 黄卡特[4] 徐炳森[2] 叶碧绿[2] 

机构地区：[1]温州医学院 [2]温州医学院附属第一医院生殖医学中心 [3]温州医学院组织胚胎学教研室 [4]温州医学院附属第一医院病理科,温州325000

出　　处：《生殖医学杂志》2007年第1期24-28,共5页Journal of Reproductive Medicine

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

摘　　要：目的探求冻融人胎儿卵巢组织异种移植后的生长发育能力。方法胎儿卵巢组织分为新鲜胎儿卵巢组织(新鲜组),慢冻-快融胎儿卵巢组织(冻融组),解冻的胎儿卵巢组织异种移植到8只裸鼠肾被膜下(移植组)三组。2个月后开始隔天注射孕马血清促性腺激素(PMSG),持续5个月。注射人绒毛膜促性腺激素(hCG)36 h后裸鼠处死取得移植物。胎儿卵巢组织石蜡包埋后连续切片,进行组织学评价和增殖细胞核抗原(PCNA)的检测。结果冻融组和新鲜组中绝大多数是原始卵泡。与新鲜组相比,冻融组中正常及总的原始卵泡数虽有下降,异常原始卵泡数虽有升高,但均无显著性差异。移植组的原始卵泡数明显少于新鲜组和冻融组(7.2±1.64,20.4±5.68,17.2±4.27,P<0.05),但可见到数个形态正常的初级卵泡、次级卵泡和窦卵泡。三组的原始卵泡均未见有PCNA阳性表达,PCNA阳性表达见于移植组初级卵泡的颗粒细胞和卵母细胞、次级卵泡和窦卵泡的颗粒细胞。结论冻融人胎儿卵巢组织异种移植后能存活,卵泡能正常发育到窦卵泡。Objective： To investigate the viability and development of follicles in the frozen-thawed human fetal ovarian tissues after xenografted. Methods： The fetal ovarian tissues were divided into three groups： fresh group, thawed group, and grafted group. The fresh fetal ovarian tissues were as fresh group, and the fetal ovarian tissue slowly cryopreserved and quickly thawed as thawed group. In the grafted group, the frozen-thawed tissues were xenografted into kidney capsules of eight nude mice. Two months later, the nude mice were injected with pregnant mare ＇s serum gonadotropin （PMSG） every other day for five months, and hCG was injected 36 hours before killing. The grafted ovarian samples were obtained and embedded in paraffin. Continuous sections of 5 μm in thickness were prepared for histological and PCNA （proliferating cell nuclear antigen） evaluation. Results： Most primordial follicles were found in the fresh group and the thawed group. Although the number of the primordial follicles with normal appearance and the total number of primordial follicles in thawed group were fewer than those in fresh group, no significant difference was found between these two groups. There were fewer primordial follicles in grafted group than those in the fresh and the thawed groups （7.2±1.64 vs 20.4±5.68 and 17.2±4.27, P〈0.05）, but many normal primary, second follicles and three antral follicles were seen in the grafted group. There was no positive PCNA expression in primordial follicles of these three groups. The positive PCNA was found in the granular cells and oocytes of primary follicles, the granular cells of secondary and antral follicles of the grafts. Conclusion. The frozen-thawed human fetal ovarian tissues could survive and develop to the stage of antral follicles successfully when xenografted.

关 键 词：卵巢组织 冷冻 异种移植 增殖 

分 类 号：R617[医药卫生—外科学]