title: Establishment and characterization of a fin cell line from blunt snout bream, Megalobrama amblycephala.
abstract: This study established and characterized a new cell line (MAF) from the fin of blunt snout bream (Megalobrama amblycephala), a freshwater fish cultivated in China. MAF cells proliferated well in medium 199 supplemented with 10 % fetal bovine serum at 28 °C and have been subcultured more than 95 times in almost a year. MAF cells were revived at 90-95 % viability after 3-6 months of storage in liquid nitrogen. Karyotyping indicated that the modal chromosome number of MAF cells was 48. The MAF cell line consisted predominantly of fibroblastic and epithelial-like cells from M. amblycephala, which was confirmed by immunofluorescence and mitochondrial 12s rRNA sequencing. Viral susceptibility tests showed that MAF cells were susceptible to infection by snakehead rhabdovirus, spring viremia carp virus, and channel catfish virus, which was demonstrated by the presence of cytopathic effect, high viral titers, and PCR products. Bacterial cytotoxicity studies showed that extracellular products from Aeromonas hydrophila were toxic to MAF cells. Cu²⁺ was also cytotoxic to MAF cells, and the 24-h IC₅₀ value was 144.48 μmol/l. When MAF cells were transfected with pEGFP-N1 plasmid, bright fluorescent signals were observed, and the transfection efficiency reached up to 5 %. These results suggest that the MAF cell line may provide a valuable tool for studying virus pathogenesis, as well as cytotoxicity testing and genetic manipulation studies.
authors.affiliation: Key Laboratory of Agricultural Animal Genetics, College of Fisheries, Breeding and Reproduction of Ministry of Education, Huazhong Agricultural University, Wuhan, 430070, People's Republic of China.
authors.forename: Dong-Mei
authors.initials: DM
authors.lastname: Zhu
authors.forename: Kun
authors.initials: K
authors.lastname: Yang
authors.forename: Wei-Min
authors.initials: WM
authors.lastname: Wang
authors.forename: Wen
authors.initials: W
authors.lastname: Song
mesh_terms: D016980:Aeromonas hydrophila; D058500:Animal Fins / Q000166:cytology*; D000818:Animals; D018929:Cell Culture Techniques; D002460:Cell Line / Q000166:cytology* / Q000187:drug effects / Q000821:virology; D003530:Cyprinidae / Q000033:anatomy & histology* / Q000821:virology; D003588:Cytopathogenic Effect, Viral; D004272:DNA, Mitochondrial; D019216:Metals, Heavy / Q000633:toxicity; D018613:Microscopy, Confocal; D013696:Temperature; D018675:Toxicity Tests
chemical_list: D004272:DNA, Mitochondrial; D019216:Metals, Heavy