Liu X F, Wu Y H, Wei S N, Wang N, Li Y Z, Zhang N W, Li P F, Qin Q W, Chen S L
Yellow Sea Fisheries Research Institute, CAFS, Key Laboratory for Sustainable Development of Marine Fisheries, Ministry of Agriculture, Qingdao 266071, China.
Laboratory for Marine Fisheries Science and Food Production Processes, Qingdao National Laboratory for Marine Science and Technology, Qingdao 266235, China.
J Fish Biol. 2018 Feb;92(2):298-307. doi: 10.1111/jfb.13471. Epub 2018 Jan 14.
A new brain-cell line, EMB, was developed from kelp grouper Epinephelus moara, a cultured marine fish. The EMB cells were subcultured for more than 60 passages. The cells were cultured in Leibovitz's L-15 medium (L15) supplemented with antibiotics, foetal bovine serum (FBS), 2-mercaptoethanol (2-ME) and basic fibroblast growth factor (bFGF). The cells could grow at 18-30° C, with the maximum growth between 24 and 30° C. The optimum FBS concentration for the cells growth ranged between 15 and 20%. Chromosome analysis indicated that the modal chromosome number was 48 in the cells at passage 45. After being transfected with pEGFP-N3 plasmid, the cells could successfully express green fluorescence protein (GFP), implying that this cell line can be used for transgenic studies. A significant cytopathic effect (CPE) was observed in the cells after infection with Singapore grouper iridovirus (SGIV) or red spotted grouper nervous necrosis virus (RGNNV) and the viral replication was confirmed by quantitative real-time PCR (qrt-PCR) assay, which suggested EMB's application potential for studies of SGIV and RGNNV.
一种新的脑细胞系EMB是从养殖海水鱼斜带石斑鱼（Epinephelus moara）中培育出来的。EMB细胞传代培养超过60代。细胞在添加了抗生素、胎牛血清（FBS）、2-巯基乙醇（2-ME）和碱性成纤维细胞生长因子（bFGF）的Leibovitz's L-15培养基（L15）中培养。细胞能在18-30°C下生长，在24-30°C时生长最快。细胞生长的最佳FBS浓度范围为15%-20%。染色体分析表明，第45代细胞的众数染色体数为48条。用pEGFP-N3质粒转染后，细胞能成功表达绿色荧光蛋白（GFP），这意味着该细胞系可用于转基因研究。用新加坡石斑鱼虹彩病毒（SGIV）或红斑石斑鱼神经坏死病毒（RGNNV）感染细胞后，观察到明显的细胞病变效应（CPE），并通过定量实时PCR（qrt-PCR）检测证实了病毒复制，这表明EMB在SGIV和RGNNV研究中的应用潜力。
