College of Animal Science and Technology, Jilin Agricultural University, Key Laboratory of Animal Production and Safety of Ministry of Education, Changchun, 130118, China.
In Vitro Cell Dev Biol Anim. 2013 Oct;49(9):734-42. doi: 10.1007/s11626-013-9661-x. Epub 2013 Jul 27.
The explosive Koi herpesvirus (KHV) epidemic has caused the deaths of a large number of carp and carp variants and has produced serious economic losses. The mirror carp (Cyprinus carpio var. specularis) exhibits strong environmental adaptability and its primary cells can be used to isolate KHV. This study utilized the tissue explant method to systematically investigate primary cell culture conditions for mirror carp snout and caudal fin tissues. We demonstrated that cells from these two tissue types had strong adaptability, and when cultured in Medium 199 (M199) containing 20% serum at 26 to 30°C, the cells from the snout and caudal fin tissues exhibited the fastest egress and proliferation. Inoculation of these two cell types with KHV-infected fish kidney tissues produced typical cytopathic effects; additionally, identification by electron microscopy, and PCR indicated that KHV could be isolated from both cell types.
爆发性锦鲤疱疹病毒(KHV)疫情已导致大量鲤鱼和鲤鱼变种死亡,并造成严重的经济损失。镜鲤(Cyprinus carpio var. specularis)表现出很强的环境适应性,其原代细胞可用于分离 KHV。本研究利用组织外植块法系统地研究了镜鲤吻部和尾部组织的原代细胞培养条件。结果表明,这两种组织来源的细胞具有很强的适应性,在含 20%血清的 M199 培养基中于 26 到 30°C 培养时,吻部和尾部组织来源的细胞具有最快的逸出和增殖速度。将这两种细胞类型接种感染 KHV 的鱼肾组织,可产生典型的致细胞病变效应;此外,通过电子显微镜和 PCR 鉴定,可从这两种细胞类型中分离到 KHV。