Establishment and Characterization of a Liver Cell Line from Spotted Knifejaw (Oplegnathus punctatus)
ZHANG Hong-Xiang1,2, LI Kun-Ming1,2, WANG Na1, ZHAI Jie-Ming3, CHEN Song-Lin1*
1 Yellow Sea Fisheries Research Institute, Chinese Academy of Fisheries Sciences, Function Laboratory for Marine Fisheries Science and Food Production Processes/Qingdao National Laboratory for Ocean Science and Technology, Qingdao 266071, China; 2 College of Fisheries and Life Sciences, Shanghai Ocean University, Shanghai 201306, China; 3 Shandong Laizhou Mingbo Aquatic Products Co., Ltd., Yantai 261400, China
Abstract：As powerful tool, fish cell lines have been widely used in virology, environmental toxicology, cell biology, genomics and genetics. Spotted knifejaw (Oplegnathus punctatus) is an excellent marine fish species. In this study, the tissue block method was used to establish the O. punctatus liver cell line (OPL) from spotted knifejaw liver. The culture medium used for the cell line was L-15 which was supplemented with mycillin, fetal bovine serum (FBS), antibiotics, basic fibroblast growth factor (bFGF) , and 2-mercaptoethanol (2-ME) . The cultured OPL cells, fibroblastic in morphology, had been subcultured to more than 30 passages in a good proliferating state. By comparing the effects of different medium of L-15 (Leibovitz-15), DMEM/F12 (Dulbecco's modified Eagle medium/Ham's F-12), DMEM and MEM on cell growth, the result showed that the optimum medium was L-15. The effects of temperature, FBS and bFGF on the growth of OPL cells were examined. The cells grew well in the temperature of 18~30 ℃, and the optimum growth temperature was 24 ℃. The results showed that the significant growth rate of OPL cells were accompanied with the increase of serum concentration range of 0%~20%. In addition, bFGF was benefit for growth of OPL cells. Chromosome analysis indicated that the passage 20 cells exhibited a normal diploid karyotype (2n=2sm+46t), but the ratio was only 68%. And the significant fluorescent signals were detected in OPL cells after transfection with pEGFP-N3 plasmid. The cytochrome oxidase subunitⅠ (COⅠ) gene was used to identified that OPL cell line originated from O. punctatus. Moreover, the establishment of the OPL cell line could provide basic experimental materials for studying the mechanisms of infective viruses and the function analysis of cells and genes.
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