MA Chen,WANG Fu-jing,LI Xia,QIN Yan-jie,LI Ya-juan
Key Laboratory of Marine Bio-resources Restoration and Habitat Reparation in Liaoning Province, Key Laboratory of Mariculture & Stock Enhancement in North China’s Sea, Ministry of Agriculture, Dalian Ocean University, Dalian 116023, China
Abstract: Primary cell culture of fin from loachMisgurnusanguillicaudatuswas carried out by tissue explant method and the cells were subcultured for 63 generations until by the special and combined aseptic processing method in which the fin tissue was immerged in 10% povidone-iodine for 15 min, and then in penicillin and streptomycin mixture (500 IU/mL penicillin and 500 μg/mL streptomycin) for 30 min. The medium DMEM/F12 used in the primary culture and early subculture was supplemented with 20% fetal bovine serum(FBS),10 ng/mL basic fibroblast growth factor(bFGF), 20 ng/mL insulin-like growth factor-I(IGF-Ⅰ)and 10 μg/mL chondroitin sulfate(pH 7.2). Only 20% FBS-DMEM/F12 was used as medium after 30 generations, and the fin cell line was cultured under the conditions of 25 ℃ with 5% CO2for 56.85 h, with chromosome number of 50 in the samples collection from 100 metaphase plates. The cell livability of(81.31%±1.46%)was fund after thawing in 30 day frozen cells. The virus sensitivity test revealed that the cell line was susceptible to the infection of spring viremia of carp virus (SVCV), with viral titer of about 105.62TCID50/mL, and not to susceptible to fish nodavirus. Establishment of fin cell line from loach makes an increase in kinds of the fish cell lines, and provides the foundation for the research of toxicology and virology.
2014, Vol. 29 
