七日龄小鼠生精小管及睾丸的冷冻保存  

Cryopreservation of Seminiferous Tubules and Whole Testes of Seven-day-old Mice

作　　者：李莲军[1] 李卫真[1] 尹革芬[1] 龚伟[1] 何志云[1] 

机构地区：[1]云南农业大学动物科学技术学院,云南昆明650201

出　　处：《云南农业大学学报》2005年第2期262-264,共3页Journal of Yunnan Agricultural University

基　　金：云南省自然科学基金(2003C0046M)

摘　　要：在DMEM培养液中添加10%小牛血清(NBS)及10%二甲基亚砜(DMSO)作为冻存液,慢速降温冷冻,液氮保存7日龄小鼠生精小管及完整睾丸,37℃水浴复苏,0 25%胰蛋白酶消化成单细胞,台盼蓝染色测定细胞复苏率。结果:生精小管及完整睾丸冷冻复苏后细胞复苏率分别为87 3%及85 0%,两复苏率之间无显著差异,与生精小管单细胞对照组相比也无显著差异。冷冻损失率分别为10 1%及12 4%。结果表明,对于7日龄小鼠生精小管及完整睾丸,以10%DMSO为抗冻剂,慢速降温冷冻,液氮保存,37℃水浴复苏是一种适宜的冷冻保存方法。Seminiferous tubules and whole testes of 7-day-old mice were slowly frozen in cryopreservative solution composing of 10% dimethylsulphoxide,10% neoborn bull serum and 80% DMEM medium, stored in liquid nitrogen, thawed in a 37℃water bath,and the recoveries were measured by trypan blue exclusion staining after testicular cells dissociated by 0.25% trypsin. The recoveries of seminiferous tubules and whole testes of 7-day-old mice were 87.3% and 85.0%, respectively. There was no significant difference between the two recoveries, and also showed no significant differences compared with single cell control. The cell loss rates after freezing/thawing were 10.1% and 12.4%, respectively. The results demonstrate that a two-step slowly freezing procedure is a suitable procedure for cryopreservation of 7-day-old mouse seminiferous tubules and whole testes in 10% DMSO freezing solution.

关 键 词：冷冻保存 生精小管 睾丸 小鼠 

分 类 号：S852.16[农业科学—基础兽医学]