Establishment and characterization of fin cell lines from redfin puffer Takifugu rubripes
WANG Jin-yun1， LI Xia1,2， QIN Yan-jie1， LI Zhuang-zhuang1， ZHANG Xiao-yu1， LIANG Yan1,JIANG Zhi-qiang2
1.Key Laboratory of Marine Bio-resource Restoration and Habitat Reparation in Liaoning Province, Dalian Ocean University, Dalian 116023, China;2.Key Laboratory of Mariculture and Stock Enhancement in North China’s Sea, Ministry of Agriculture and Rural Affairs, Dalian Ocean University, Dalian 116023, China
Abstract: A tissue block culture method is adopted to initiate primary culture of fin tissue cells of redfin pufferTakifugurubripes, and the trypsin digestion method is used for subculture. At present, the fin cells of redfin puffer have passed to 40 generations and become the fibroblast-like cells. The medium used for primary culture and early subculture was DMEM/F12 medium supplemented with 20% fetal bovine serum, with basic fibroblast growth factor(bFGF) final concentration of 10 ng/mL, type I insulin-like growth factor of 20 ng/mL(IGF-1), and the chondroitin sulfate final concentration of 20 μg/mL. DMEM/F12 medium containing 20% fetal bovine serum was used, and cells were cultured in a constant temperature incubator at 25 ℃ and 5% CO2 in the 30th generation. The growth curve showed that the cells were in the incubation period from 0 to 2 days, in the logarithmic growth period from 2 to 4 days, in the plateau growth period from 4 to 5 days, and in the recession period 5 days afterwards. The culture doubling time was found to be 50.24 h in redfin puffer fin cell line with survival rate of 80%, in 60 days cryopreservation, after thawing and resuscitation. Microsatellite site amplification and gel electrophoresis revealed that the cell line was originated from redfin puffer. The establishment of the redfin puffer fin cell line enriches the species of fish cell lines and lays a foundation for the research of viral diseases and vaccine preparation in redfin puffer.
2018, Vol. 33 
