四种抗冻剂20℃时对小鼠生精上皮单细胞存活率的影响  

Effects of Four Cryoprotectants on Survival of Single Cells from Mouse Seminiferous Epithelia at 20 ℃

作　　者：李莲军[1] 施江滨[1] 陆峻波[1] 

机构地区：[1]云南农业大学动物科技学院,云南昆明650201

出　　处：《动物医学进展》2002年第2期54-56,共3页Progress In Veterinary Medicine

摘　　要：将小鼠生精上皮单细胞置 2 0℃含不同抗冻剂的冻存液中一定时间后 ,台盼蓝染色测定细胞存活率 ,以筛选冷冻保存小鼠生精上皮细胞的侯选抗冻剂及使用浓度。结果 ,2 0℃ 3 0min,1 0 %浓度的二甲基亚砜 ( DMSO)、丙二醇( PG)及乙二醇 ( EG)对 7日龄小鼠生精上皮单细胞存活率均无显著影响 ,而 1 0 %甘油 ( G)则使细胞存活率显著下降 ;2 0℃ 3 0 min,1 0 %浓度的 DMSO、PG、EG及 G对成年小鼠生精上皮单细胞存活率均无显著影响 ;2 0℃ 5min,2 5%浓度的 DMSO、PG、EG及 G均使 7日龄及成年小鼠生精上皮单细胞存活率显著下降。实验结果表明 ,1 0 % DMSO、PG、及 EG可作为 7日龄小鼠生殖细胞慢速冷冻保存时的侯选抗冻剂 ,1 0 % DMSO、PG、EG及 G可作为成年小鼠生殖细胞慢速冷冻保存时的侯选抗冻剂 ;在高浓度抗冻剂超速冷冻保存小鼠生殖细胞时 ,平衡时间应短于 5min,或在 4℃进行。The survival rates of single cells from mouse seminiferous epithelia were measured by trypan blue exclusion after a definite time span in various cryopreservative solutions at 20 ℃. Dimethyl sulphoxide (DMSO), propanediol(PG) or ethylene glycol(EG) at concentration of 10% at 20 ℃ all show no significant different effects on the survival rates of 7 day old mouse seminiferous epithelial cells compared with control, while glycerol(G) significantly decreased the survival rates. DMSO, PG, EG and G at concentration of 10% all show no significant different effects on the survival rates of adult mouse male germ cells compared with control. DMSO, PG, EG and G at concentration of 25% all significantly decreased the survival rates of both 7 day old mouse seminiferous epithelial cells and adult mouse male germ cells. The results indicate that DMSO, PG and EG are likely cryoprotectants for slow cryopreservation of 7 day old mouse seminiferous epithelial cells. DMSO, PG, EG and G at concentration of 10% are likely cryoprotectants for slow cryopreservation of adult mouse male germ cells. When mouse seminiferous epithelial cells are ultrarapidly cryopreserved in a cryopeservative solution at a high concentration, the balance time should be shorter than 5 min, or operate at 4 ℃.

关 键 词：抗冻剂 生精上皮 生殖细胞 睾丸 小鼠 冷冻保存 存活率 

分 类 号：S814.3[农业科学—畜牧学] R321.1[农业科学—畜牧兽医]