【摘要】 研究了冷冻保护液、平衡时间、降温速度、解冻液以及解冻方式对冷冻小鼠胎儿成纤维细胞和牛睾丸成纤维细胞活力的影响。结果表明 ,对小鼠胎儿成纤维细胞和犊牛睾丸成纤维细胞进行冷冻 ,冷冻液 ( 0 .4 %BSA) +1 .5mol/ L乙二醇 +0 .1 mol/ L蔗糖的 PBS液 )的冷冻效果优于冷冻液 ( 1 0 %的 NBS+1 0 %DMSO的 DMEM液 )。用冷冻液 对小鼠胎儿成纤维细胞和犊牛睾丸成纤维细胞进行冷冻 ,平衡时间可从 2 .5h缩短到 0 .5h;冷冻细管降温速度 ( -6.8～ -3 7℃ )可以从 0 .6℃ / min提高到 1 .2℃ / min;诱发结晶可以省略。 3种解冻液解冻细胞活力排序为 :解冻液 ( 0 .4 %BSA+0 .1 mol/ L蔗糖的 PBS) >解冻液 ( 1 0 %NBS的 DMEM) >解冻液 ( PBS)。用冷冻液 对 MEF和 BTF细胞冷冻过程中 ,无需用培养液制备细胞悬液反复离心更多还原

【Abstract】 Cryopreservation of murine embryo fibroblast and new bovine testicular fibroblast were investigated in this paper.The results showed as follows: The vitality of murine embryo fibroblast(MEF) cells and new bovine tisticular fibroblast(NBTF) cells exposed to the freezing media Ⅱ(PBS supplemented 0.4% BSA,1.5 mol/L ethylene glycol(EG),0.1 mol/L sucrose) higher than freezing media Ⅰ(10% NBS+10%DMSO+DMEM); When NBTF and MEF were freezed in media Ⅱ,the balanced time can be reduced from 2.5 h to 0.5 h without influence on their vitality. Moreover,freezing rate can be increased from 0.6℃/min to 1.2℃/min,induced ice did not influence on their vitality.On the vitality of NBTF cells and MEF cells,thawing media Ⅱ(PBS supplemented 0.4% BSA,0.1 mol/L sucrose)>thawing media Ⅲ(DMEM supplemented 10% NBS)>thawing media ⅠPBS.更多还原