莎能奶山羊皮肤成纤维细胞几种冷冻保存方法的研究

以莎能奶山羊皮肤组织为材料进行细胞培养,通过分离纯化建立了莎能奶山羊皮肤成纤维细胞系.纯化培养的成纤维细胞在冷冻保护剂和血清成分相同的条件下选用5种具有不同预冷平衡方式和预冷时间的方法进行冷冻保存,5个月后通过对成纤维细胞复苏后的活力对比分析,方法3(70%细胞悬液 20%胎牛血清 10%DMSO;先在4 C预冷平衡0.5 h,接着液氮罐口气态氮悬挂4 h,然后沉入液氮)冻存细胞解冻后胎盘蓝染色细胞活力分析,活细胞率为84.8%,培养48 h后细胞贴壁率为85.4%,明显高于其他几种方法.

Analyses Research of Several Cryopreservation Methods for Skin Fibroblasts of Shaneng Milk Goat

Through isolation and pure culture of Shaneng milk goat fibroblasts with skin tissue, fibroblast line was established. On conditions of the same cryoprotectant and FCS concentration Pure Shaneng milk goat fibroblasts cultured were cryopreservated in five cryopreservation methods with different pre-cool and balance ways and pre-cool time, through the contrast and analyses of cell vigor after five months cryopreservation, method 3 (70% cell suspension 20?S 10% DMSO; cryopreservation procedure: first fibroblasts were pre-cooled and balanced for half hour at 4 C , and second hanged in gaseous nitrogen in close to the mouth of nitrogen canister for 4 hours, and finally sunk into liquid nitrogen. ) was the best cryopreservation method, it could maintain 84. 8% of living cell rate through cell vigor test after cells were revived and stained by Trypan Blue and 85. 4% of cell adherence rate after cells were revived and cultured for 48 hours, living cell rate and cell adherence rate of method 3 superior to other four methods.