成年小鼠生精小管及睾丸的冷冻保存  

Cryopreservation of Adult Mouse Seminiferous Tubules and Whole Testes

作　　者：李莲军[1] 何志云[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》2005年第6期72-74,共3页Progress In Veterinary Medicine

基　　金：云南省自然科学基金(2003C0046M)

摘　　要：采用含不同浓度抗冻剂二甲基亚砜(DMSO)、乙二醇(EG)、丙二醇(PG)及丙三醇(G)的DMEM培养基作为冻存液,对成年小鼠生精小管及完整睾丸进行慢速冷冻,液氮保存,37 ℃水浴复苏,并测定细胞复苏率。结果表明,成年小鼠生精小管在50 mL/L、100 mL/L 及150 mL/L DMSO冻存液中的细胞复苏率分别为92.6%、93.4%及69.4%,在50 mL/L、100 mL/L及150 mL/L PG 冻存液中的细胞复苏率分别为94.2%,93.3%及66. 9%,在50 mL/L、100 mL/L 及150mL/L EG 冻存液中的细胞复苏率分别为87.2%、91. 0%及62. 6%,在50 mL/L、100mL/L及150 mL/L G冻存液中的细胞复苏率分别为90.0%、90.5%及67.1%。各种抗冻剂的最高细胞复苏率之间差异不显著(P> 0. 05 )。成鼠完整睾丸在含100 mL/LDMSO、PG、EG或G的冻存液中的细胞复苏率分别为56.0%、50.0%、50.2%及50.2%。结果显示,DMSO、PG、EG 及G 均适宜用作成年小鼠生精小管的抗冻剂,最佳使用浓度均为50 mL/L^100 mL/L。完整睾丸冷冻后的细胞复苏率低于60%,仍需进一步研究。Adult mouse seminiferous tubules and whole testes were slowly frozen in different freezing solutions, stored in liquid nitrogen, thawed in a 37 ℃ water bath, and the cell recovery rates were examined by trypan blue exclusion staining after thawing, respectively. The freezing solutions were DMEM 100 g/L NBS supplemented with 50 mL/L, 100 mL/L, or 150 mL/L dimethyl sulphoxide (DMSO), propanediol(PG), ethylene glycol(EG), or glycerol(G),respectively. The cell recoveries of adult mouse seminiferous tubules in freezing solutions with DMSO at concentrations of 50 mL/L, 100 mL/L and 150 mL/L were 92.6%, 93.2%, and 69.4%, respectively. The cell recoveries in freezing solutions with PG at concentrations of 50 mL/L, 100 mL/L and 150 mL/L were 94.2%, 93.3%, and 66.9%, respectively. The cell recoveries in freezing solutions with EG at concentrations of 50 mL/L, 100 mL/L and 150 mL/L were 87.2%, 91.0%, and 62.6%, respectively. The cell recoveries in freezing solutions with G at concentrations of 5%, 10% and 15% were 90.0%, 90.5%, and 67.1%, respectively. There were no significant differences among the highest recoveries of four cryoprotectants. The results suggest that four cryoprotectaots all are suitable for cryopreservation of adult mouse seminiferous tubules, and their favorable concentration all are at 50 mL/L^100 mL/L. The cell recoveries of adult mouse testes are all below 60%, and further research is needed.

关 键 词：冷冻保存 抗冻剂 生精小管 睾丸 小鼠 

分 类 号：S857.29[农业科学—临床兽医学]