In order to screen the optimal culture system for Vero cell culture, this study has optimized the culture medium, passage ratio and density for three consecutive passage respectively. Above all, Vero cells were cultured in different kinds of medium for three passages to select the best medium. Subsequently, different subculture density and proportion of Vero cells were cultured in the optimal medium for three passage, and the optimized passage ratio win out. In general, 1.296% MEM medium supplied with superfine fetal bovine serum is better than the other combination of medium and it can sustain the successive stable culture passaging. In the condition of this medium, the best proportion of cell passaging is 1∶6 and the optimal cell density is 3×10⁴/cm². In this study, we screened the conditions of the best Vero cell culture system and confirmed that the conditions meet the continuous and stable passage of Vero cells.