Abstract:Based on single-factor experiments，response surface methodology was employed to optimize the cultivation process of HepG2 cellsin roller bottles.The optimal culture conditions were found that 4×107HepG2 cells were incubated each bottle cultured for 48 h with rotate speed of 9 r/min.The predicted HepG2 cells yield under these conditions was 10.56×107cells/bottle and experimental value was 10.88×107cells/bottle with the relative error being about 3.03%.The results in vitro antitumor experiments showed that seleno-chitosan could significantly inhibit the proliferation of HepG2 cells in 96-well plate and roller bottles with IC50of 169 μg/mL and 245 μg/mL respectively，suggesting that the HepG2 cells cultured in roller bottles would cause cells aggregation and generate drug antagonists.The experimental results provide a theoretical basis for development of cancer vaccine and drug treatment of tumor.