State Key Laboratory of Freshwater Ecology and Biotechnology, Institute of Hydrobiology, Chinese Academy of Sciences, Wuhan 430072, China.
Huaian Fisheries Scientific Research Institute of Jiangsu Province, Huaian 223001, China.
Virus Res. 2014 Aug 30;189:56-62. doi: 10.1016/j.virusres.2014.04.013. Epub 2014 Apr 30.
An Ussuri catfish Pseudobagrus ussuriensis skin (UCS) cell line was developed and subcultured for more than 60 passages. UCS cells consisted of mostly epithelial-like cells and multiplied well in TC199 medium supplemented with 10% fetal bovine serum at 25°C. Chromosome analysis revealed that most UCS cells had a normal diploid karyotype with 2n=52. UCS cells showed differential cytopathic effects (CPEs) after inoculation of spring viremia of carp virus (SVCV, a negative-strand RNA virus), grass carp reovirus (GCRV, a multi-segmented double-stranded RNA virus) and Rana grylio virus (RGV, a large double-stranded DNA virus), and were indicative of high sensitivities to these three aquatic animal viruses by a virus titration study. The CPE caused by SVCV appeared as rounded and granular cells, grape-like clusters and small lytic plaques. Characteristic CPE containing plaque-like syncytia was induced by GCRV. RGV-infected cells produced typical CPE characterized by cells shrinkage and aggregation, formation of clear plaques and cell sheet detachment. Furthermore, significant fluorescent signals were observed after UCS cells were transfected with green fluorescent protein reporter plasmids, and the development of CPE induced by a recombinant RGV, ΔTK-RGV, in UCS cells was illustrated using a combination of light and fluorescence microscopy. The data from this study suggested that UCS cell line can potentially serve as a useful tool for the comparison study of different aquatic animal viruses and the isolation of some newly emerging viruses in Ussuri catfish farming.
乌苏里拟鲿皮肤（UCS）细胞系的建立与传代培养超过 60 代。UCS 细胞主要由上皮样细胞组成，在添加 10%胎牛血清的 TC199 培养基中于 25°C 下良好增殖。染色体分析显示，大多数 UCS 细胞具有正常的二倍体核型，2n=52。UCS 细胞在接种鲤鱼春病毒血症病毒（SVCV，负链 RNA 病毒）、草鱼出血病病毒（GCRV，多节双链 RNA 病毒）和牛蛙病毒（RGV，大型双链 DNA 病毒）后出现明显的细胞病变效应（CPE），通过病毒滴定研究表明其对这三种水生动物病毒高度敏感。SVCV 引起的 CPE 表现为圆形和颗粒状细胞、葡萄状簇和小溶斑。GCRV 诱导产生具有合胞体特征的典型 CPE。RGV 感染的细胞产生典型的 CPE，表现为细胞皱缩和聚集、形成清晰的蚀斑和细胞单层脱落。此外，UCS 细胞转染绿色荧光蛋白报告质粒后，观察到明显的荧光信号，并用荧光显微镜和明场显微镜观察了重组 RGVΔTK-RGV 在 UCS 细胞中诱导的 CPE 发展情况。该研究数据表明，UCS 细胞系可能成为比较不同水生动物病毒和乌苏里拟鲿养殖中某些新出现病毒的有用工具。
