Wei S, Yu Y, Qin Q
Department of Aquatic Animal Medicine, College of Marine Sciences, South China Agricultural University, Guangzhou, 510642, China.
Laboratory for Marine Biology and Biotechnology, Qingdao National Laboratory for Marine Science and Technology, Qingdao, 266071, China.
J Fish Biol. 2018 Jun;92(6):1675-1686. doi: 10.1111/jfb.13566. Epub 2018 Apr 24.
A new cell line derived from the caudal fin of golden pompano Trachinotus ovatus (TOCF) was successfully established and characterized. TOCF cells grew well at 28° C in L-15 medium supplemented with 10% foetal bovine serum (FBS). The cell line has been subcultured in more than 100 passages. Molecular characterization of 18S ribosomal (r)RNA and cytochrome oxidase subunit 1 (COI) confirmed that the TOCF cells were derived indeed from T. ovatus. TOCF cells have a modal chromosome number of 54. It was further showed that TOCF cells were transfected successfully with pEGFP-N3 and pDsRED-N1 plasmid, suggesting that TOCF cells could be used to research gene functions in vitro. Viral susceptibility tests showed that TOCF cells were susceptible to Singapore grouper iridovirus (SGIV), observed by the occurrence of the cytopathic effect (CPE) with the formation of inclusion bodies. In addition, the expression of major capsid protein (MCP) gene of SGIV changed during virus infection in TOCF cells. Thus, our present results described the characteristic of a TOCF cell line that could be a valuable tool for genetic manipulation, as well as isolation and propagation of iridovirus studies.
成功建立并鉴定了一种源自卵形鲳鲹尾鳍的新细胞系（TOCF）。TOCF细胞在添加10%胎牛血清（FBS）的L-15培养基中于28°C生长良好。该细胞系已传代培养超过100次。18S核糖体（r）RNA和细胞色素氧化酶亚基1（COI）的分子特征证实TOCF细胞确实源自卵形鲳鲹。TOCF细胞的染色体众数为54。进一步表明，pEGFP-N3和pDsRED-N1质粒成功转染了TOCF细胞，这表明TOCF细胞可用于体外基因功能研究。病毒敏感性试验表明，通过出现细胞病变效应（CPE）并形成包涵体观察到，TOCF细胞对新加坡石斑鱼虹彩病毒（SGIV）敏感。此外，SGIV主要衣壳蛋白（MCP）基因在TOCF细胞病毒感染过程中的表达发生了变化。因此，我们目前的结果描述了一种TOCF细胞系的特征，该细胞系可能是基因操作以及虹彩病毒研究的分离和增殖的宝贵工具。
