Establishment And Characterization Of A Cell Line Derived From Fin Of Paddlefish, Polyodon Spathula Walbaum

Along with the rapid development of sturgeons' artificially farming industry in the aspects of constant increase in cultural scpecies, farming scale and the intensitivity, the problems of sturgeon diseases have been serious more than ever before. The outbreak of diseases in cultured sturgeons happened very frequently and caused huge economic losses. Fish cell lines are the most fundamental and important materials for the studies on isolation and identification of viral pathogens of aquatic animals, pathogenic biology, mechanism of infection, diagnostic technology, vaccine preparation and immunoprophylaxis, etc. At present, several pathogenic viruses of sturgeons have been reported at abroad by using the established cell lines derived from the sturgeons, but at home only few preliminary studies on sturgeon cell cultivation have been conducted. There is a desperate shortage of cell lines derived from sturgeons currently. Paddlefish, polyodon spathula Walbaum, is one of the most popular cultured species of sturgeons, to establish a cell lie derived from the tissue of paddlefish may provide a crucial material for the studies on sturgeon viral diseases and the control.By using tissue explant technique, a cell line derived from the fin of paddlefish has been established and designated as PF-Fin. The subculture of PF-Fin cell line is conducted at 25℃in M199 medium supplemented with 10% fetal bovine serum and has been constantly splited over 80 times. The cell monolayer consists of fibroblast-like cells. After 8 months cryopreservation of PF-Fin cells in liquid nitrogen, PF-Fin cells exhibit a viability of 87.68±3.61% by typan blue stained.Characterizing PF-Fin cells growth with different medium (M199, MEM, R1640, L15), fetal bovine serum concentration (0%,5%,10%,20%) and at different temperature (15℃,20℃,25℃,30℃), the results show that the optimal conditions for PF-Fin cell line are M199 media,10% fetal bovine serum concentration and 25℃. The plating efficiency of PF-Fin cells is about 13.75±2.28%. Chromosome typing analysis reveals that the chromosome number of PF-Fin cell line is 2n=120 at the 5th passage, and at its 59th passage the chromosome modal number is 2n=90. The chromosome number of PF-Fin cell line has changed significantly. The sensitivity tests of viral infections indicate that PF-Fin cell line is susceptible to the infection of Grass crap Reovirus (GCRV), Channel catfish Reovirus (CCRV) and Spring Viremia of carp Virus (SVCV) and a characteristic viral-induced cytopathic effect (CPE) is observed. Application of PF-Fin cell line in isolating sturgeons'viral pathogens is currently going on.