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Establishment and characterization of a mid-kidney cell line derived from golden pompano Trachinotus ovatus, a new cell model for virus pathogenesis and toxicology studies

机译:Establishment and characterization of a mid-kidney cell line derived from golden pompano Trachinotus ovatus, a new cell model for virus pathogenesis and toxicology studies

摘要

Golden pompano Trachinotus ovatus, a popularly cultured and commercially important marine fish worldwide, has been recognized as a promising candidate for mariculture. However, outbreaks of infectious bacterial or viral diseases and environmental deterioration have led to great economic losses in T. ovatus aquaculture recently. In our research, we established a new mid-kidney cell line, designated as TOK, from golden pompano, T. ovatus. The optimized growth temperature and working concentration of fetal bovine serum (FBS) were 28A degrees C and 10-20%, respectively. Foreign genes could express well in TOK cells. The modal number of TOK cells was 54. The TOK cells were susceptive to Singapore grouper iridovirus (SGIV) and red-spotted grouper nervous necrosis virus (RGNNV), and the virus could propagate in cells. Propagation was verified by qRT-PCR, and virions were observed under electron microscopy. Cytotoxicity analysis revealed that TOK cells were sensitive to different concentrations of extracellular products (ECPs) from Vibrio alginolyticus and V. anguillarum. Moreover, heavy metals (Cd, Cu, and Hg) also showed dose-dependent cytotoxicity to the TOK cell line. We established a mid-kidney cell line from T. ovatus which could be applied to cytotoxicity assays of heavy metals. The newly established TOK cell line possesses great application potential in genetic manipulation, virus-host interaction studies, and toxicity assays of bacterial extracellular products and heavy metals.
机译:金po鱼Trachinotus ovatus是一种在世界范围内具有广泛养殖和商业意义的海鱼,已被公认为是海水养殖的有前途的候选人。但是,感染性细菌或病毒性疾病的暴发以及环境的恶化最近导致卵形水产养殖的巨大经济损失。在我们的研究中,我们建立了一个新的中肾细胞系,命名为TOK,它来自金色po参(T. ovatus)。胎牛血清(FBS)的最佳生长温度和工作浓度分别为28A摄氏度和10-20%。外源基因可以在TOK细胞中良好表达。 TOK细胞的模式数为54。TOK细胞易受新加坡石斑鱼虹膜病毒(SGIV)和红斑石斑鱼神经坏死病毒(RGNNV)的感染,并且该病毒可以在细胞中传播。通过qRT-PCR验证了繁殖,并且在电子显微镜下观察到病毒体。细胞毒性分析表明,TOK细胞对不同浓度的溶藻弧菌和鳗弧菌细胞外产物(ECP)敏感。此外,重金属(Cd,Cu和Hg)对TOK细胞系也表现出剂量依赖性的细胞毒性。我们建立了一种来自卵圆线虫的中肾细胞系,该细胞系可用于重金属的细胞毒性测定。新近建立的TOK细胞系在基因操作,病毒-宿主相互作用研究以及细菌细胞外产物和重金属的毒性测定中具有巨大的应用潜力。

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