SCC-3737℃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℃ and its application in common virus culture

作　　者：巩金鹏 赵颖 陈雪兰 蔺凌云[1] 姚嘉赟[1] 穆雪姣 周可欣 王丛旭 沈锦玉[1,3] 潘晓艺[1] GONG Jinpeng;ZHAO Ying;CHEN Xuelan;LIN Lingyun;YAO Jiayun;MU Xuejiao;ZHOU Kexin;WANG Congxu;SHEN Jinyu;PAN Xiaoyi(Key Laboratory of Fish Health and Nutrition of Zhejiang Province,Key Laboratory of Healthy Freshwater Aquaculture,Ministry of Agriculture and Rural Affairs,Zhejiang Institute of Freshwater Fisheries,Huzhou 313001,China;Huzhou Nanxun Centre for Agricultural Technology Extension and Service,Huzhou 313009,China;College of Fisheries and Life Science,Shanghai Ocean University,Shanghai 201306,China)

机构地区：[1]浙江省淡水水产研究所,农业农村部淡水渔业健康养殖重点实验室,浙江省鱼类健康与营养重点实验室,浙江湖州313001 [2]湖州市南浔区农业技术推广服务中心,浙江湖州313009 [3]上海海洋大学水产与生命学院,上海201306

出　　处：《水产学报》2025年第3期233-242,共10页Journal of Fisheries of China

基　　金：国家重点研发计划(2019YFD0900103);浙江省自然科学基金(ZCLTGN24C1901);浙江省“三农九方”科技协作计划(2024SNJF053)。

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℃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103.4~109.6TCID50/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。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℃.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 103.4 to 109.6 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.

关 键 词：鳜 鳜脊髓组织细胞系 37℃驯化 生长特性 敏感性 

分 类 号：S943[农业科学—水产养殖]