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中国鲈鱼(Siniperca chuatsi)皮肤细胞系的建立与特性鉴定及其在水产动物病毒研究中的应用。

Development and characterization of a skin cell line from Chinese perch (Siniperca chuatsi) and its application in aquatic animal viruses.

机构信息

Institute of Hydrobiology, Chinese Academy of Sciences, Wuhan, China.

College of Modern Agriculture Sciences, University of the Chinese Academy of Sciences, Beijing, China.

出版信息

J Fish Dis. 2022 Oct;45(10):1439-1449. doi: 10.1111/jfd.13673. Epub 2022 Jun 28.

Abstract

Chinese perch (Siniperca chuatsi), an important fish for the aquaculture industry of China, is often affected by viral diseases. A stable and sensitive cell line can play an important role in virus identification and isolation, functional gene identification, virus pathogenic mechanism and antiviral immunity study. In the present study, a new cell line (S. chuatsi skin cell, SCSC) derived from the skin of S. chuatsi was established. The SCSC mainly consisted of fibroblastic-like cells, which grew well in M199 medium supplemented with 10% foetal bovine serum at 25°C. Chromosome analysis revealed that the SCSC (44%) has a diploid chromosome number of 2n = 48. The SCSC can be transfected and expressed exogenous gene efficiently. It also showed high sensitivity to several aquatic animal viruses from different families including Rhabdoviridae, Iridoviridae and Reoviridae. In addition, RT-PCR showed that S. chuatsi rhabdovirus (SCRV) started genome replication as early as 3 h post infection in the cells, which also induced the up-regulation of a variety of immune-related genes including these related to interleukin family, pattern recognition receptors, JAK-STAT pathway and interferon regulatory factors. In summary, current study provided a new tool in research of fish viruses and its interaction with host.

摘要

中国鲈鱼(Siniperca chuatsi)是中国水产养殖业的重要鱼类,常受到病毒病的影响。稳定而敏感的细胞系在病毒鉴定和分离、功能基因鉴定、病毒致病机制和抗病毒免疫研究中发挥着重要作用。本研究从中国鲈鱼的皮肤中建立了一种新的细胞系(S. chuatsi 皮肤细胞,SCSC)。SCSC 主要由成纤维样细胞组成,在 25°C 下,M199 培养基中添加 10%胎牛血清时生长良好。染色体分析表明,SCSC(44%)具有二倍体染色体数 2n=48。SCSC 可有效转染和表达外源基因。它对来自不同科的几种水生动物病毒也表现出高度敏感性,包括弹状病毒科、虹彩病毒科和呼肠孤病毒科。此外,RT-PCR 显示,S. chuatsi 弹状病毒(SCRV)在感染后 3 小时内就开始在细胞中复制基因组,这也诱导了多种免疫相关基因的上调,包括与白细胞介素家族、模式识别受体、JAK-STAT 通路和干扰素调节因子相关的基因。总之,本研究为鱼类病毒及其与宿主的相互作用研究提供了一种新工具。

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