Establishment and characterization of a new cell line derived from half-smooth tongue sole (Cynoglossus semilaevis) kidney
Establishment and characterization of a new cell line derived from half-smooth tongue sole (Cynoglossus semilaevis) kidney
复制标题
半滑舌鳎 (Cynoglossus semilaevis) 肾脏来源的新细胞系的建立和表征
DOI：
10.1007/s00343-019-9084-y
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发表时间：
2019
影响因子：
1.6
通讯作者：
肖鹏
中科院分区：
文献类型：
--
作者：
娄雅楠;孙斌;张丽娜;李勇;肖鹏
本研究从半滑舌鳎（Cynoglossus semilaeviskidney cells，CSK）的肾脏中建立了一个新的细胞系。传代培养1000 d以上，传代100次以上。此外，将CSK细胞最佳维持在补充有HEPES、抗生素、胎牛血清、2-巯基乙醇和碱性成纤维细胞生长因子的Dulbecco改良Eagle培养基营养混合物F-12中。CSK细胞的最适生长温度为25°C，并且细胞显示成纤维细胞样表型。染色体分析显示CSK细胞具有正常的二倍体核型，2n=42。石斑鱼神经坏死病毒（NNV）感染CSK细胞后3- 5d即可引起细胞病变。CSK细胞的NNV敏感性与细胞质中病毒粒子的高丰度有关，如通过电子显微镜观察到的。此外，CSK细胞可以成功地转染一个绿色荧光蛋白报告质粒，并很容易观察到荧光信号。免疫细胞化学分析显示CSK细胞为支持细胞。总体而言，我们建立了这个新的细胞系，这可能有潜在的应用在鉴定病毒病原体影响半光滑舌的唯一。
In this study, we established a new cell line (Cynoglossus semilaeviskidney cells, CSK) from kidney of the half-smooth tongue sole (Cynoglossus semilaevis). The cells were subcultured over 1 000 days and passaged for more than 100 times. Additionally, CSK cells were optimally maintained in Dulbecco’s modified Eagle medium nutrient mixture F-12 supplemented with HEPES, antibiotics, fetal bovine serum, 2-mercaptoethanol, and basic fibroblast growth factor. The optimum growth temperature for CSK cells was 25°C, and the cells showed a fibroblast-like phenotype. Chromosome analysis revealed that CSK cells had a normal diploid karyotype with 2n=42. CSK cells were susceptible to Grouper nervous necrosis virus (NNV), and cytopathic effects were observed at 3–5 days postinfection. The NNV sensitivity of CSK cells was related to the high abundance of virions in the cytoplasm, as observed by electron microscopy. Additionally, CSK cells could be successfully transfected with a green fluorescent protein reporter plasmid, and fluorescent signals were easily observed. Finally, immunocytochemistry analysis showed that CSK cells were supporting cells. Overall, we established this new cell line, which may have potential applications in the identification of viral pathogens affecting the half-smooth tongue sole.