SCC-37 37 °C驯化的稳定性及其在常见病毒培养中的应用

Study on the stability of SCC-37 acclimated at 37 °C and its application in common virus culture

摘要:

目的 建立能快速增殖且对鳜源常见病毒敏感的鳜鱼细胞系。方法 利用已建立的鳜脊髓组织细胞系SCC，采用梯度血清逐步驯化的方式，建立一个连续细胞系SCC-37，分析该连续细胞系的来源、支原体污染、生长特性和冻存后细胞活力，评估其对传染性脾肾坏死病毒(ISKNV)、鳜虹彩病毒(SCRaV)和鳜弹状病毒(SCRV)的敏感性。结果 该细胞系SCC-37已连续培养30代，稳定的生长条件为L-15+10%NBCS、37 °C培养；通过鳜28S rRNA基因分析其来源于鳜，支原体检测证实SCC-37无支原体污染；病毒敏感性结果显示，ISKNV、SCRaV和SCRV可在SCC-37中稳定增殖和传代，培养的病毒滴度为10^3.4~10^9.6 TCID₅₀/mL，电镜观察可见大量病毒颗粒。结论 新驯化的SCC-37为分离、鉴定和增殖病毒以及疫苗制备提供了重要工具。

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.