Cell culture and plaque forming test of five arboviruses

Object: To select a sensitive cell and to establish the best cell culture conditions used in studying biological characters of five arboviruses. Methods: Suckling mouse brain suspensions at different dilutions were inoculated to monolayer of BHK_(21), LLC-MK_2, A_(549) and C6/36 cells to test their sensitivity to the viruses. Cytopathic effect(CPE) appearing time, virus titers and character of CPE were also observed. Then, the L_9(3~3) orthogonal design test of BHK_(21) cell culture and plaque forming test were conducted. Results: All the five arboviruses could be cultured in the four cell strains and all caused earlier appearance of CPE in BHK_(21) cells which showed distinct CPE character and higher titer than in the other three cell strains. The result of orthogonal test of five arboviruses in culturing cells showed plaques in BHK_(21) cell monolayer. Conclusions: Under the condition of 60% DMEM, 30% hydrolyzed lactalbumin(HL)(0.5%), 10% serum, 100 U/ml penicillin and streptomycin and 50 ug/ml kanamycin, pH7.2, BHK_(21) cell could grow well enough to fulfil the requirement of studying biological characters of five arboviruses.