Development and characterization of a new tropical marine fish cell line from grouper, Epinephelus coioides susceptible to iridovirus and nodavirus
Development and characterization of a new tropical marine fish cell line from grouper, Epinephelus coioides susceptible to iridovirus and nodavirus
复制标题
DOI：
10.1016/j.jviromet.2005.07.009
复制
发表时间：
2006-01-01
影响因子：
3.1
通讯作者：
Zhang, RQ
中科院分区：
文献类型：
--
作者：
Qin, QW;Wu, TH;Zhang, RQ
本文报道了一种新的热带海洋鱼类细胞系（GS）的建立和鉴定。GS是从橙子斑点石斑鱼（Epinealopus coioides）脾脏中分离得到的。GS细胞在添加10%胎牛血清的Leibovitz's L-15培养基中生长良好，并已传代200多次。最适生长温度为27 ℃。GS细胞培养物主要由成纤维细胞组成。二倍体众数为48。GS细胞培养物在感染致病性石斑鱼虹彩病毒（新加坡石斑鱼虹彩病毒，SGIV）或石斑鱼诺达病毒（石斑鱼神经坏死病毒，ETNNV）后显示出晚期细胞病变效应。通过透射电子显微镜分析显示，在病毒感染的细胞的细胞质中分别存在大量的SGIV和ETNNV颗粒，表明对这两种病毒的高敏感性。免疫荧光显微镜显示，感染虹彩病毒的GS细胞与抗石斑鱼虹彩病毒的单克隆抗体反应强烈。这表明GS细胞系具有良好的潜力，作为诊断工具的分离和繁殖的虹彩病毒和野田病毒。当用pEGFP载体DNA转染GS细胞时，观察到显著的荧光信号，表明GS细胞系可用作转基因和遗传操作研究的有用工具。(c)2005 Elsevier B. V.保留所有权利。
The development and characterization of a new tropical marine fish cell line (GS), derived from the spleen of orange spotted grouper, Epinephelus coioides is described. The GS cells grow well in Leibovitz's L-15 medium supplemented with 10% foetal bovine serum, and have been subcultured more than 200 times. The optimal growth temperature was 27 degrees C. The GS cell culture consisted of mostly fibroblastic cells. The modal diploid chromosome number was 48. GS cell cultures showed advanced cytopathic effects after infection with a pathogenic grouper iridovirus (Singapore grouper iridovirus, SGIV) or with a grouper nodavirus (Epinephelus tauvina nervous necrosis virus, ETNNV). Analysis by transmission electron microscopy showed a large number of SGIV and ETNNV particles in the cytoplasm of virus-infected cells, respectively, indicative of high sensitivity to these two viruses. Immunofluorescence microscopy showed that iridovirus-infected GS cells reacted strongly with monoclonal antibody against the grouper iridovirus. It is suggested that the GS cell line has good potential as a diagnostic tool for isolation and propagation of iridovirus and nodavirus. When the GS cells were transfected with pEGFP vector DNA, significant fluorescent signals were observed suggesting that the GS cell line can be used as a useful tool for transgenic and genetic manipulation studies. (c) 2005 Elsevier B.V. All rights reserved.