小鼠胎儿成纤维细胞的冷冻保存研究

Study on the cryopreservation of mouse fetus fibroblast cells

为了摸索小鼠胎儿成纤维细胞冷冻保存的简易方法，采用常规冷冻和玻璃化冷冻2种模式，探讨了不同的冷冻方法、不同的冷冻前平衡时间、不同的冷冻保护液的配比等因素对成纤维细胞冷冻效果的影响。实验结果表明：常规冷冻采用含100,4（体积分数）甘油的DMEM液作保护剂，可将0～5℃下平衡时间缩短至45～60min，细胞活力可达0．87。玻璃化冷冻采用10％（体积分数）NBS＋1．8mol／L乙二醇＋0．1mol／L蔗糖的PBS冷冻小鼠胎儿成纤维细胞效果好于其他组合，细胞活力可达到0．86。玻璃化冷冻法与常规冷冻法的冷冻效果差异不显著。

In order to get facility method for the cryopreservation of mouse fetus fibroblast cells, we adopted convention freezing method and vitrification method to study the effect of different freezing methods,different banlance time, and different ratio of component in the protective medium. The result showed that the banlance time at 0-5 ℃ could be reduced to 45-60 min and the vitality be 0.87 when we took 10% glycerol＋DMEM as the cryoprotector. For the mouse fetus fibroblast cells,the best vitrification method is taking 10%NBS＋ 1.8 mol/L glycol＋0. 1 mol/L sucrose as cryoprotector, and the vitality can be 0. 86. Vitrification and convention freezing method had not significant difference.