Abstract:Abstract Fish cells are important tools for studying virology, immunology, genetics, and so on. Turbot (Scophthalmus maximus) is a widely cultivated marine fish species in Europe and China. However, no brain cell line has been developed so far. S. maximus brain cell line (SMB) was established by tissue block method from turbot brain in this research. The cell line was maintained in DMEM/F12 which was supplemented with 4-(2-hydroxyethyl)-1-piperazineethanesulfonic acid (Hepes), antibiotic (mycillin), fetal bovine serum (FBS), antibiotics, basic fibroblast growth factor (bFGF), and 2-mercaptoethanol (2-ME). The cultured SMB cells were composed of fibroblast-like cells and epithelial-like cells, and they had been subcultured for more than 40 passages. The cells grew well in the temperature 18~30 ℃, and the optimum growth temperature was 24 ℃. Chromosome analysis indicated that the passage 30 cells exhibited chromosomal aneuploidy, and 43% of the cells maintained a normal diploid chromosome number (2n=44t). When the SMB cells were successfully transfected with pEGFP-N3 plasmid, the transfection efficiency was up to about 30%. The SMB cell line will provide a new important experimental material to study the mechanisms of infective viruses and gene function analysis.