Establishment And Charactetizaton Of Kidney Cell Lines From Rough Skin Sculpin (Trachidermus Fasciatus)

This paper described the establishment of kidney cell line from rough skin sculpin(Trachidermus fasciatus) and research the biological characterization.The results showed that the explant method was suitable to this tissue.The results showed that the kidney cells migrated from their tissues after cultured in vitro for 36 h. The kidney cells in primary culture were all in fibroblastic morphology and proliferated to confluence within 30 d. All the cells were subcultured by trypsin digestion method. They grew at a steady rate during subsequent subcultures. After passage 20, kidney cells could be subcultured after 5-6d. Kidney cells have been subcultured up to passage 55. Continuous rough skin sculpin kidney cell line have been successfully established. The doubling time were 56.32 h in kidney cells at the 35 th passage. Rough skin sculpin had a diploid chromosome number of 40 and its karyotype formula was 2n= 40= 14m+10sm+16t. The result of virus sensitivity test indicated that the cell line is susceptible to the infection of SVCV,a characteristic viral-induced cytopathic efect(CPE)was observed and the viral titer reached about 106.53TCID50/m L。The kidney cells were cultured in different medium fetal bovine serum concentration(0%,5%,10%,15%,20%),different chondroitin sulfate concentration(0、10、20、30、40μg/m L), different Human basic fibroblast growth factor concentration(0、2.5、5、7.5、10ng/m L) and different the type I insulin-like growth factor concentration(0、10、20、30ng/m L). The results show that the optimal conditions for the kidney cell line are 15% fetal bovine serum concentration, 20μg/m L chondroitin sulfate concentration, 20ng/m L the type I insulin-like growth factor concentration and 7.5ng/m L Human basic fibroblast growth factor concentration.This study researched on the toxic effects of glyphosate on kidney cells, the 24 h semi-lethal concentration of kidney cells ere1177.66mg/L respectively, which were measured with MTT method. Micronucleus test results show that there have no visible difference between the experimental group and the control group.