正交试验法优选HepG2细胞最佳培养条件

Optimization of the culture conditions of HepG2 cells by orthogonal design

目的优选HepG2细胞最佳培养条件。方法分别以培养基、细胞接种密度、血清含量、双抗浓度、pH、Hanks清洗次数、胰酶浓度和消化时间为量化指标设计正交试验,SPSS11.0统计软件分析,得方差分析及多重比较结果。结果含15%胎牛血清的DMEM培养基,接种密度为4×104个/mL,双抗浓度为0.5%,pH为7.2;在汇合度达95～100%时,Hanks洗3次,0.05%胰酶-EDTA-Na2H2O消化1min,传代效果最好,条件易于控制,且细胞能顺利生长。结论本法为HepG2细胞最佳培养条件的确定提供了可靠的实验数据。

[Objective] To optimize the best cuhure condition of HepG2. [Method] Design the orthogonal test taking culture medium, inoculative density, serum content, penicillin and streptomycin concentration, pH, the times of washing by Hanks＇ , trypsin concentration and digest time as the factors. Obtain the variance analysis and multicomparison results by the statistical software SPSS 11.0. [Result] Cells growed well in DMEM that contain 15% fetal cattle serum, and inoculative density was 4×10^4 cell/ml, penicillin and streptomycin concentration was 0.05%, pH was 7.2. It was easy to control and have good digest effect that use Hanks washes the cells three times advance then apply 0.05% trypsin-EDTA-Na2H2O to digest for one minute when convergence degree reaches 95～100%. [Conclusion] This method provide credible experimental datum for confirming the best culture condition of HepG2.