首页> 外文期刊>Journal of Fish Biology >A novel fish cell line derived from the brain of Chinese perch Siniperca chuatsi: development and characterization
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A novel fish cell line derived from the brain of Chinese perch Siniperca chuatsi: development and characterization

机译:一种来自中国鲈鱼Siniperca chuatsi的大脑的新型鱼细胞系:发育与鉴定

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摘要

In this study, a continuous cell line (named as CPB) was established from Siniperca chuatsi brain and has been subcultured >140 times. CPB cell line predominantly consisted of fibroblast-like cells that could grow better in Leibovitz's L-15 supplemented with 10% foetal bovine serum at 28 degrees C. Polymerase chain reaction amplification of 18s recombinant (r)RNA confirmed the origin of this cell line from S.chuatsi. The CPB cell line was cryopreserved at different passage levels and revived successfully with 80-90% survival. The cell line was further characterized by chromosome number and transfection. The CPB cells were highly susceptible to infectious spleen and kidney necrosis virus (ISKNV) with a titre of 658-662 log TCID50 ml(-1) and numerous ISKNV particles were observed in the cytoplasm by transmission electron microscopy. At the same time, ISKNV infection was confirmed by reverse transcriptase polymerase chain reaction, immunodot blot and individual challenge experiments. The development and characterization of a new brain cell line from S. chuatsi were described in this study and it could be used as an in vitro tool for propagation of ISKNV and gene expression studies.
机译:在这项研究中,从Siniperca chuatsi脑中建立了一个连续的细胞系(命名为CPB),并进行了140次以上的传代培养。 CPB细胞系主要由成纤维细胞样细胞组成,这些细胞可以在Leibovitz的L-15中补充28%的10%胎牛血清,从而生长更好。18s重组(r)RNA的聚合酶链反应扩增证实了该细胞系起源于沙棘CPB细胞系在不同的传代水平下冷冻保存,并成功恢复了80-90%的存活率。该细胞系通过染色体数目和转染进一步表征。 CPB细胞高度敏感,感染滴度为658-662 log TCID50 ml(-1)的脾脏和肾脏坏死病毒(ISKNV),通过透射电子显微镜在细胞质中观察到许多ISKNV颗粒。同时,通过逆转录聚合酶链反应,免疫斑点印迹和单独的攻击实验证实了ISKNV感染。在这项研究中描述了来自沙特氏菌的一种新的脑细胞系的发展和特征,它可以用作ISKNV繁殖和基因表达研究的体外工具。

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