Hepa1-6细胞的最佳培养条件研究

Study on the Optimum Culture Conditions of Hepa1-6 Cells

[目的]探索Hepa1-6细胞的最佳培养条件。[方法]以小鼠Hepa1-6肝癌细胞为材料,采用正交试验设计研究RPMI1640和DMEM2种不同培养基及细胞接种密度、血清浓度、双抗浓度、D-Hanks漂洗次数、胰蛋白酶浓度和消化时间6个不同因素对Hepa1-6传代培养的影响;根据细胞在6个时间点的生长状况评分,筛选出Hepa1-6细胞的最佳培养条件。[结果]Hepa1-6细胞在含有20%血清的RPMI-1640培养基,接种密度为4×104个/cm2,100U/ml的双抗浓度,贴壁率在90%以上时,D-Hanks漂洗1次,0.5%胰蛋白酶消化2min,传代效果最好。[结论]通过正交设计筛选,为Hepa1-6细胞体外培养探索出最佳培养条件。

[Objective]The research aimed to explore the optimum culture conditions of Hepa1-6 cells.[Method] Hepa1-6 hepatoma cell lines of mice were used to make the orthogonal test.The effects of RPMI1640 and DMEM media,cell inoculation density,serum concentrations,penicillin and streptomycin concentration,the times of washing by D-Hanks,digestion time and concentration of trypsin on the passage culture of Hepa1-6 cells were studied.Based on the growth situations of cells at 6 time points,the optimum culture conditions of Hepa1-6 cells were screened out.[Result]Hepa1-6 cells grew well in the RPMI-1640 medium that contained 20% serum,and inoculation density was 4×104 cell/cm2,penicillin and streptomycin concentration of 100 U/ml.It’s have good digest effect that D-Hanks was used to wash the cells once,0.5% trypsin was digested for 2 min when the convergence rate was more than 90% .[Conclusion]Through the orthogonal design,the optimum culture conditions of Hepa1-6 cells in vitro were explored.