Objective To study the culture characteristics of tick-borne encephalitis virus (TBEV) in susceptible cells and identify the RNA replication, protein expression and infectivity.Methods The TBEV was cultured using African green monkey kidney cells (Vero cells). The RNA replication of TBEV was detected by quantitative polymerase chain reaction, the expression of flavivirus envelope protein was detected by Western blotting, and the titer of TBEV was detected by plaque assay. The human lung adenocarcinoma cells (A549 cells) was infected with TBEV, and the cytopathic effect of A549 cells infected with TBEV was observed under microscope. The expression of flavivirus envelope protein in A549 and Vero cells infected with TBEV was detected by immunofluorescence staining.Results The levels of TBEV RNA were increased in Vero cells at 72 h incubation compared with those at 48 h incubation, and the expression of flavivirus envelope protein was detectable. The results of plaque assay showed that the TBEV titer in the supernatant of Vero cells was (2.0±1.4)×10⁶ plaque-forming unit (PFU)/mL after culturing for 72 h. There was obvious cytopathic effect in A549 cells with TBEV infection. The positive rate of flavivirus envelope protein expression in A549 cells (61.0%) was higher than that in Vero cells (9.3%) after infection with 1∶1 000-diluted TBEV. Compared with Vero cells, the titer in the supernatant of A549 cells infected with TBEV for 48 h was significantly higher ([2.0±0.4]×10⁷ PFU/mL vs [8.5±2.1]×10³ PFU/mL, P < 0.05).Conclusion Vero cells can be used to culture TBEV, and A549 cells are more susceptible to TBEV.