Establishment and characterization of a fin cell line from blunt snout bream, Megalobrama amblycephala

被引:0
作者
Dong-Mei Zhu
Kun Yang
Wei-Min Wang
Wen Song
机构
[1] Breeding and Reproduction of Ministry of Education,Key Laboratory of Agricultural Animal Genetics, College of Fisheries
[2] Huazhong Agricultural University,undefined
来源
Fish Physiology and Biochemistry | 2013年 / 39卷
关键词
Fin; Cell line; Cytotoxicity; Transfection;
D O I
暂无
中图分类号
学科分类号
摘要
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.
引用
收藏
页码:1399 / 1410
页数:11
相关论文
共 260 条
[1]  
Babu VS(2011)Establishment and characterization of a fin cell line from Indian walking catfish, J Fish Dis 34 355-364
[2]  
Nambi KSN(2001) (L.) Zool Res 22 502-506
[3]  
Chandra V(1995)Molecular cloning and sequence analysis of insulin-like growth factor-I cDNA from bluntnose bream ( Microbiologia 11 439-446
[4]  
Ishaq Ahmed VP(1997)) Aquaculture 150 143-153
[5]  
Bhonde R(1980)Role of extracellular products in the pathogenicity of Vibrio strains on cultured gilt-head seabream ( In Vitro 16 365-368
[6]  
Sahul Hameed AS(2013)) Arch Environ Contam Toxicol 64 301-315
[7]  
Bai JJ(1999)A continuous cell line from the cultured marine fish gilt-head seabream ( J Fish Dis 22 173-182
[8]  
Lao HH(2009) L.) J Huazhong Agr Univ 28 604-608
[9]  
Ye X(2010)Fish cell lines establishment of a line from ovaries of channel catfish Biochim Biophys Acta 1801 526-536
[10]  
Li YH(1975)Effects of waterborne chronic copper exposure on hepatic lipid metabolism and metal-element composition in Methods Cell Sci 1 31-35