人脐带间充质干细胞移植于不育小鼠睾丸的研究  

Study on Transplantation of Human Umbilical Cord Mesenchymal Stem Cells into Infertile Mouse Testis

作　　者：唐秋灵[1] 林广裕[1] 林丽敏[1] 黄天华[2] 谢庆东[2] 马廉[1] 

机构地区：[1]汕头大学医学院第二附属医院儿科 [2]汕头大学医学院生殖医学研究中心,广东汕头515041

出　　处：《实用儿科临床杂志》2009年第23期1796-1799,1805,共5页Journal of Applied Clinical Pediatrics

基　　金：国家自然科学基金项目资助(30772354);广东省社会发展计划项目资助(2007B080701022)

摘　　要：目的观察人脐带华尔通胶来源的间充质干细胞(MSCs)移植于不育小鼠睾丸的克隆能力及对其生殖功能的影响。方法采用腹腔内单次注射白消安的方法构建不育小鼠模型;采用组织块贴壁法获得MSCs,分别用Brdu和Hoechst33258标记,显微注射法移植入不育小鼠睾丸的精曲小管内;采取免疫组织化学和快速冷冻切片荧光显微镜下观察的方法检测其移植细胞存活情况;采用HMIAS-2000高清晰度彩色医学图文分析系统测量小鼠睾丸精曲小管直径。应用SPSS13.0软件进行统计学分析。结果白消安单次注射可造成15%的小鼠死亡,用药后第4周,不育小鼠睾丸的精曲小管仅存基底膜,呈薄的空管状;传至3代即可得到数量较多的纯化MSCs,MSCs移植于异体小鼠睾丸获得成功,移植后的细胞能存活至少4个月,并且发生了从管腔到基底膜再到管腔的移行过程;移植后26d与120d,不育模型移植细胞组精曲小管直径均较未移植细胞组有所增加,且其差异有统计学意义(P<0.01)。结论采用精曲小管显微注射法将MSCs移植于不育小鼠睾丸获得成功,移植后的MSCs能长期存活并发生移行,且能促进不育小鼠生殖功能的恢复。Objective To observe clone ability of human umbilical cord mesenchymal stem cells （MSCs） into infertile mouse seminife-rous tubules and the effects of MSCs on reproductive function.Methods Busulfan was used to destroy endogenous spermatogenesis of the recipient mice.To isolate,culture and purify MSCs with adherent method before marked with Brdu and Hoechst 33258 respectively,and then transplanted into the seminiferous tubules by microinjection.The survival of MSCs in recipient testes were evaluated by immunohistochemistry stained for Brdu and Fluorescent microscopy for Hoechst 33258 observation at different times.The diameter of seminiferous tubules was detected with HMIAS-2000 high-definition colored analyzing system for medical pictures.SPSS 13.0 software was used to analyze the data.Results The dosage of Busulfan resulted in 15% death in the mice,the testis of survived mice showed only basilar membrane in seminiferous tubules after 4 weeks.A lot of purified MSCs were obtained at the third generation and transplantation them into mouse seminiferous tubules survive for at least 4 months and appear to migration.The average diameter in experimental groups were higher than those in controls not only on 26 days but also on 120 days（P〈0.01）.Conclusions Transplantation of MSCs using mice as recipients is feasible and effective.The MSCs after transplantation not only survive for a long time and appear to migration but also play an energetic role in improving the reproductive function.

关 键 词：脐带 间充质干细胞 睾丸 移植 生殖功能 

分 类 号：R698.2[医药卫生—泌尿科学]