Jing Hongli, Gao Longying, Zhang Min, Wang Na, Lin Xiangmei, Zhang Lifeng, Wu Shaoqiang
Research Centre of Aquatic Animal Diseases, Institute of Animal Quarantine, Chinese Academy of Inspection and Quarantine, Beijing, 100029, China.
Shekou in Shenzhen Entry-Exit Inspection and Quarantine Bureau, Shenzhen, 518067, China.
Fish Physiol Biochem. 2016 Feb;42(1):303-11. doi: 10.1007/s10695-015-0138-6. Epub 2015 Sep 29.
Goldfish Carassius auratus are commonly used in scientific research and have a significant economic value in the pet trade. In this study, two cell lines were established from the snout and kidney tissue of goldfish, in order to create a biological monitoring tool for viral diseases. Cell lines were optimally maintained at 25 °C in M199 medium supplemented with 15-20 % fetal bovine serum. Chromosomal analysis indicated that both cell lines remained diploid, with a mean chromosomal count of 100. Results of viral inoculation assays revealed that both cell lines shared similar patterns of viral susceptibility and production to infectious pancreatic necrosis virus (IPNV). The viral titers of IPNV in goldfish snout cell line (GFSe) and goldfish kidney cell line (GFKf) reached 10(6.8) and 10(5.9) TCID50/0.1 mL, respectively, within 7 days. The cytopathic effect could be observed when Cyprinid herpesvirus 2 was inoculated into these cells; however, the CPE disappeared after four passages. No CPE was observed in the cells infected by spring viremia carp virus or grass carp reovirus. These newly established cell lines will be a useful diagnostic tool for viral diseases in this fish species and also for the isolation and study of goldfish viruses in future.
金鱼(Carassius auratus)常用于科学研究,在宠物贸易中具有重要的经济价值。在本研究中,从金鱼的吻部和肾脏组织建立了两个细胞系,以创建一种针对病毒性疾病的生物监测工具。细胞系在补充有15 - 20%胎牛血清的M199培养基中于25°C下最佳培养。染色体分析表明,这两个细胞系均保持二倍体状态,平均染色体数为100。病毒接种试验结果显示,这两个细胞系对传染性胰腺坏死病毒(IPNV)的病毒易感性和产生模式相似。在7天内,金鱼吻部细胞系(GFSe)和金鱼肾脏细胞系(GFKf)中IPNV的病毒滴度分别达到10(6.8)和10(5.9) TCID50/0.1 mL。将鲤疱疹病毒2接种到这些细胞中时可观察到细胞病变效应;然而,传代四次后细胞病变效应消失。在感染鲤春病毒血症病毒或草鱼呼肠孤病毒的细胞中未观察到细胞病变效应。这些新建立的细胞系将成为该鱼类病毒性疾病的有用诊断工具,也将用于未来金鱼病毒的分离和研究。