冷冻保存未成熟睾丸组织移植后生精细胞的生长发育  

Spermatogenesis of cryopreserved immature mouse testicular tissues being allografted into immunodeficient mice

作　　者：于洁[1] 万汇涓[1] 尹美珺[1] 叶静[1] 张芳婷[1] 肖琴[1] 房家智[1] 

机构地区：[1]北京大学深圳医院中心实验室,深圳518036

出　　处：《中国男科学杂志》2008年第12期15-18,共4页Chinese Journal of Andrology

基　　金：广东省自然科学基金资助项目(No.04007312);深圳市科技计划项目(No.200404101No.200803004)

摘　　要：目的采用睾丸组织移植模型,探讨通过简单易行的冷冻保存程序保存未成熟睾丸组织,并使冷冻后的组织恢复生精过程的可能性,为该动物模型应用在保存雄性生殖细胞、帮助癌症患者保存生殖能力以及保存濒危物种等方面提供进一步的实验依据。方法用DMEM培养基加入二甲基亚砜配制冷冻保存液,将新生1～2d昆明小鼠睾丸组织按分步冷冻的方法进行冷冻,最终放入-80℃超低温冰箱中保存起来。2周后取出冻存的睾丸组织,移植到雄性去势免疫缺陷小鼠背部皮下,分别于移植后4周、5周和8周取材,制作HE染色切片,观察移植物中生精小管结构和生精细胞的组成,并与新鲜移植的睾丸组织进行对比分析。结果采用实验中的冷冻程序保存的新生小鼠睾丸组织,在冷冻保存一段时间后再移植,其表现与新鲜睾丸组织移植相同,其中末成熟的生精细胞可以在受体中继续生长发育,并完成整个生精过程,发育成为精子。结论本实验中所采用的睾丸组织冷冻保存方法具有不需特殊设备、简便易行、适用范围广泛等优点,适用于多种场合中生精细胞的冷冻保存。Objective To investigate spermatogenesis of cryopreserved immature mouse testicular tissues using animal model of testicular allografting. Methods DMEM medium supplemented with DSMO was prepared as cryopreserved solution. Testicular tissues of neonatal Kunming mice （1-2d） were gradually cooled using refrigerator and then stored at -80~C for use. Two weeks later for cryopreservation, testicular tissues were grafted into castrated male immunodeficient mice. After 4wk, 5wk and 8wk, graft samples were collected for histochemistry analysis of the development of testicular tissues and the composition of seminiferous epithelia. Results The spermatogenetic activity of the cryopreserved immature testicular tissues maintained after cryopreservation some time which was proved by the spermatogenesis obersevation. The spermatogenetic activity of the cryopreserved immature testicular tissues was similar with fresh grafted testicular tissues. Conclusion Protocol for cryopreservation was simple and easily performed in the study which could be applied widely for cryopreservation of testicular tissues in different research.

关 键 词：睾丸 低温保存 小鼠 睾丸组织移植 

分 类 号：Q954.61[生物学—动物学]