In this study, the technology for the large-scale cultivation of gibel carp brain cells and CyHV-2 by Cephodex microcarrier in suspension system was investigated. The results showed that the Cephodex microcarrier is suitable for the growth of anchorage-dependent GiCB cells. The optimal serum concentration for cells attaching to the Cephodex microcarrier was 10%, the microcarrier concentration was 6 g/L and the density of cells inoculated was 2.5×10⁵ cells/mL. With an intermittent agitation for 2 min at 35 rpm after 30 min stilling culture, the attachment efficacy reached 90% after 8 h cultivation; while in growing period, the optimal speed was 45 r/min for continuous agitation. After infection of GiCB cells on Cephodex microcarrier with CyHV-2 at a multiplicity of infection (MOI) of 0.2, the typical cytopathic effect (CPE) appeared after 5 days post-infection and the virus titer (TCID₅₀/mL) reached 10^6.50±0.30. This study established a solid basis for the large-scale preparation of vaccine against the crucian carp hematopoietic necrosis.