Guo C Y, Huang Y H, Wei S N, Ouyang Z L, Yan Y, Huang X H, Qin Q W
Key Laboratory of Tropical Marine Bio-resources and Ecology, South China Sea Institute of Oceanology, Chinese Academy of Sciences, Guangzhou, China; University of Chinese Academy of Sciences, Beijing, China.
J Fish Dis. 2015 Feb;38(2):175-86. doi: 10.1111/jfd.12221. Epub 2013 Dec 26.
A new marine fish cell line, derived from the heart of giant grouper, Epinephelus lanceolatus (Bloch), was established and characterized. The cell line was designated as ELGH and subcultured with more than 60 passages. The ELGH cells were mainly composed of fibroblast-like cells and multiplied well in Leibovitz's L-15 medium supplemented with 10% foetal bovine serum (FBS) at 28 °C. Chromosome analysis indicated that the modal chromosome number was 48. The fluorescent signals were detected in ELGH when transfected with green fluorescent protein reporter plasmids. The 50% cytotoxic concentration (CC50 ) of the extracellular products (ECPs) from Streptococcus iniae and Vibrio alginolyticus E333 on ELGH cells was 60.02 and 12.49 μg mL(-1), respectively. Moreover, the ELGH cells showed susceptibility to Singapore grouper iridovirus (SGIV), but not to soft-shelled turtle iridovirus (STIV), red-spotted grouper nervous necrosis virus (RGNNV) and spring viremia of carp virus (SVCV), which was demonstrated by the presence of a severe cytopathic effect (CPE) and increased viral titres. In addition, electron microscopy observation showed that abundant virus particles were present in the infected cells. Taken together, our data above provided the potential utility of ELGH cells for transgenic and genetic manipulation, as well as cytotoxicity testing and virus pathogenesis.
建立并鉴定了一种源自鞍带石斑鱼（Epinephelus lanceolatus，布洛赫）心脏的新型海水鱼类细胞系。该细胞系被命名为ELGH，并已传代60多次。ELGH细胞主要由成纤维样细胞组成，在添加10%胎牛血清（FBS）的Leibovitz's L - 15培养基中于28℃下生长良好。染色体分析表明，其众数染色体数为48。用绿色荧光蛋白报告质粒转染ELGH细胞后可检测到荧光信号。海豚链球菌和溶藻弧菌E333的胞外产物（ECPs）对ELGH细胞的50%细胞毒性浓度（CC50）分别为60.02和12.49 μg mL(-1)。此外，ELGH细胞对新加坡石斑鱼虹彩病毒（SGIV）敏感，但对中华鳖虹彩病毒（STIV）、红斑石斑鱼神经坏死病毒（RGNNV）和鲤春病毒血症病毒（SVCV）不敏感，这通过严重细胞病变效应（CPE）的出现和病毒滴度的增加得以证实。另外，电子显微镜观察显示感染细胞中存在大量病毒颗粒。综上所述，我们以上的数据表明ELGH细胞在转基因和基因操作、细胞毒性测试以及病毒致病机制研究方面具有潜在应用价值。
