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Establishment and Characterization of a Cell Line Derived From Fin of Paddlefish， Polyodon spathula Walbaum
Yi Xiao1,2, Ling-Bing Zeng1,2*, Xiao-Li Li2, Yuan-Chao Zou 1, Yong Zhou 1, Jin Xu2
1College of Fisheries, Huazhong Agricultural University, Wuhan 430070, China; 2Yangtze River Fisheries Research Institute,Chinese Academy of Fishery Sciences, Jingzhou 434000, China
Abstract: A cell line， designated PF-Fin， had been established from the fin of paddlefish， Polyodon spathula Walbaum and subcultured over 80 passages. Characterization of the cell line included determination of cell morphology，
optimal growth kinetics， plating efficiency， karyotyping， etc. The primary culture of this cell was generated by the tissue explant technique using the M199 medium supplemented with 30% fetal bovine serum， epidermal/fibroblast
growth factors and penicillin/streptomycin/-amphotericin B solution. Subculture of the cell was conducted in M199 medium with 10% fetal bovine serum. The cell grew between 20~30 ºC and the optimal growth temperature was 25 ºC. The cell monolayer consisted of fibroblast-like cells and the plating efficiency of the cells was about 13.75%±2.28%. Following cryopreservation in liquid nitrogen， thawed cells exhibited a viability of 87.68%±3.61% after 8 months storage period. Chromosome typing of this cell line revealed the diploid chromosome number was 2n=120 at its 10th passage， and at its 59th passage the chromosome modal number was 2n=90. The results of virus challenge tests indicated that PF-Fin cell line is susceptible to the infection of Grass carp Reovirus (GCRV)， Channel catfish Reovirus (CCRV) and Spring Viriemia carp Virus (SVCV)， and a characteristic viral-induced cytopathic effect (CPE) was observed.
optimal growth kinetics， plating efficiency， karyotyping， etc. The primary culture of this cell was generated by the tissue explant technique using the M199 medium supplemented with 30% fetal bovine serum， epidermal/fibroblast
growth factors and penicillin/streptomycin/-amphotericin B solution. Subculture of the cell was conducted in M199 medium with 10% fetal bovine serum. The cell grew between 20~30 ºC and the optimal growth temperature was 25 ºC. The cell monolayer consisted of fibroblast-like cells and the plating efficiency of the cells was about 13.75%±2.28%. Following cryopreservation in liquid nitrogen， thawed cells exhibited a viability of 87.68%±3.61% after 8 months storage period. Chromosome typing of this cell line revealed the diploid chromosome number was 2n=120 at its 10th passage， and at its 59th passage the chromosome modal number was 2n=90. The results of virus challenge tests indicated that PF-Fin cell line is susceptible to the infection of Grass carp Reovirus (GCRV)， Channel catfish Reovirus (CCRV) and Spring Viriemia carp Virus (SVCV)， and a characteristic viral-induced cytopathic effect (CPE) was observed.
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