Splenic cell line from sea bass, Lates calcarifer: Establishment and characterization

The development and characterization of a new tropical marine fish cell line (SISS), derived from the spleen of sea bass, Lates calcarifer is described. The cell line was maintained in Leibovitz's L-15 supplemented with 15% fetal bovine serum. This cell line has been sub-cultured more than 70 times over a period of one and half years. The cells were able to grow at temperature between 25 and 32 °C with optimum temperature of 28 °C. The growth rate of sea bass spleen cells increased as the FBS proportion increased from 2 to 20% at 28 °C with optimum growth at the concentrations of 15 or 20% FBS. The SISS cell line consists of predominantly of fibroblastic and epithelial-like cells. Polymerase chain reaction products were obtained from SISS cells and tissues of sea bass with primer sets of microsatellite markers of sea bass. Five fish viruses were tested on this cell line to determine its susceptibility to these viruses and this was found to be susceptible to IPNV VR-299 and nodavirus, and the infection was confirmed by RT-PCR and CPE. Further, this cell line is characterized by immunocytochemistry, endocytotic uptake of gold nanoparticles using confocal-laser-scanning microscopy (CFLSM), transfection with pEGFP-N1, proliferate marker.