褐点石斑鱼三种组织细胞系的建立

Establishment of Three Cell Lines from Brown-Marbled Grouper,Epinephelus Fuscoguttatus

为建立褐点石斑鱼鳍、心脏和鳔组织细胞系,本文利用不同培养液和培养温度对褐点石斑鱼鳍、心脏和鳔组织细胞进行了原代培养和传代培养。实验结果显示,在24℃培养于含有羧甲基壳寡糖、碱性成纤维样生长因子、I型胰岛素样生长因子及20%胎牛血清的DMEM/F12培养液(pH=7.2)中的3种细胞,均为成纤维样细胞,其生长分裂状态最佳,可以持续稳定传代。第60代褐点石斑鱼鳍、心脏和鳔细胞的群体倍增时间分别为50.6 h、40.3 h和43.3 h,其特征性染色体数目均为48条。目前鳍、心脏和鳔细胞已分别传至第90代、第70代和第75代,已成功建立了3种细胞的连续性细胞系,为鱼类病毒与宿主细胞相互作用机制等鱼类病毒学基础研究,以及鱼类病毒的分离、鉴定、繁殖及病毒疫苗研制等奠定了基础。

To establish novel cell lines from brown-marbled grouper, Epinephelus fuscoguttatus, primary cultures of fin, heart and swim bladder tissues were initiated in this study. These cells were successfully cultured at 24 ℃ by using 20 % FBS-DMEM/F12 medium （pH = 7.2）, which was further supplemented with carboxymethyl-chitooligosaccharide, basic fibroblast growth factor and insulin-like growth factor-I. The fin, heart and swim bladder cells all showed fibroblastic morphology. These cells grew at a steady rate during subsequent subculture with population doubling times of 50.6 h, 40.3 h and 43.3 h at passage 60, respectively. Chromosome analyses show that these cells exhibited chromosomal aneuploidy, but still had a modal chromosome number of 48. To date, three novel brown-marbled grouper cell lines have been established and subcultured to more than 70 passages. These continuous cell lines may serve as valuable tools for studies of cell-virus interactions and have potential applications in fish virus isolation, propagation and vaccine development.