Abstract: The SCC-37 cell line was established via acclimation, and the impact of this process on cell characteristics was analyzed. Additionally, the sensitivity of these cells to viruses was assessed, thereby reducing culture costs and laying the groundwork for large-scale vaccine production. In this study, a continuous cell line was derived from the spinal cord tissue of mandarin fish, Siniperca chuatsi, using a gradual acclimation process with a serum gradient. The origin of the cell line, mycoplasma contamination status, growth characteristics, and viability after cryopreservation were evaluated. Furthermore, the sensitivity to infectious spleen and kidney necrosis virus (ISKNV), S. chuatsi ranavirus (SCRaV), and S. chuatsi rhabdovirus (SCRV) was examined. The cell line, designated SCC-37, has been successfully cultured for up to 30 passages in complete medium (L-15+10%NBCS) at 37 °C. The S. chuatsi origin of SCC-37 was confirmed through the S. chuatsi 28S rRNA gene. Mycoplasma testing verified that SCC-37 is free of contamination. Sensitivity experiments demonstrated that ISKNV, SCRaV, and SCRV can proliferate and be passaged stably in SCC-37, with virus titers ranging from 10^3.4 to 10^9.6 TCID₅₀/mL. Electron microscopy revealed a high abundance of virus particles in SCC-37 cells. In summary, the newly acclimated SCC-37 cell line provided a crucial tool for virus isolation, identification, and amplification, as well as for vaccine preparation. Its diverse applications merited further investigation.