Establishment and characterization of a liver cell line from silver pomfret (Pampus argenteus) for studying fish health.

Abstract

Fish cell lines have become a useful tool to study in resource conservation, genetic breeding, diseases control, and environmental pollutants detection. The silver pomfret (Pampus argenteus) is a high-valued marine fish species in aquaculture, which is seriously threatened by various fish diseases. In this study, a new cell line derived from P. argenteus liver (PaL) was established and characterized. PaL cells mainly consisted of fibroblast-like morphology and multiplied well in Leibovitz's L-15 medium supplemented with 15% foetal bovine serum and 3 ng/mL basic fibroblast growth factor at 22°C. Amplification of the Cyt b gene confirmed that the origin of PaL cells as P. argenteus. Chromosome analysis revealed that PaL cells had a diploid Karyotyp. The PaL cells were efficiently transfected with pEGFP-N3 plasmids, indicating its potential application in foreign gene manipulation studies. The PaL cells were found to be susceptible to red sea bream iridovirus (RSIV) and the expression of immune-related gene (TLR5) and apoptosis-related genes (Bax, Cyt c3, CASP9) were upregulated. Furthermore, lipopolysaccharide and palmitic acid (PA) treatments decreased cell viability and up-regulated the expression of inflammation related genes (IL-8, IL-1β). Meanwhile, PA incubation induced cell apoptosis by Bcl-2-regulated caspase activation. In conclusion, the newly established PaL cell line will be an appropriate in vitro tool for viral propagation and immune response.