Establishment,characterization, virus susceptibility and transfection of cell lines from cobia,Rachycentron canadum (L.), brain and fin |
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| Authors: | T‐C Cheng Y‐S Lai I‐Y Lin C‐P Wu S‐L Chang T‐I Chen M‐S Su |
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| Institution: | 1. Department of Tropical Agriculture and International Cooperation, National Pingtung University of Science and Technology, Pingtung, Taiwan;2. Graduate Institute of Biotechnology, Ilan University, Ilan, Taiwan;3. Department of Life Science, Mingdao University, Changhua, Taiwan;4. Department of Food Science, Ocean University, Keelung, Taiwan;5. Tungkan Biotechnology Research Center, Fisheries Research Institute, Pingtung, Taiwan;6. Fisheries Research Institute, Keelung, Taiwan |
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| Abstract: | Establishment and characterization of two cobia, Rachycentron canadum, cell lines derived from cobia brain (CB) and cobia fin (CF) are described. Caudal fin and brain from juvenile cobia were dissociated for 30 and 10 min, respectively, in phosphate‐buffered saline containing 0.25% trypsin at 25 °C. The optimal culture condition for both dissociated cells (primary cell culture) was at 28 °C in Leibovitz‐15 medium containing 10% foetal bovine serum. The cells have been sub‐cultured at a ratio of 1:2 for more than 160 passages over a period of 3 years. Origin of the cultured cells was verified by comparison of their sequences of mitochondrial cytochrome oxidase subunit I genes (cox I) with the cox 1 sequence from cobia muscle tissue. The cell lines showed polyploidy. No mycoplasma contamination was detected. Susceptibility to grouper iridovirus was observed for the CB cell line but not the CF cell line. Both cell lines expressed green fluorescent protein after being transfected with green fluorescent reporter gene driven by the cytomegalovirus promoter. |
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| Keywords: | brain cell line cobia cox I fin |
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