Development, characterization and application of a new fibroblastic-like cell line from kidney of a freshwater air breathing fish Channa striatus (Bloch, 1793)

A new cell line, Channa striatus kidney (CSK), derived from the kidney tissue of murrel, was established and characterized. The CSK cell line was maintained in Leibovitz's L-15 supplemented with 10% fetal bovine serum and has been subcultured more than 140 times. This cell line was able to grow in a range of temperatures from 22 to 32 degrees C with optimal growth at 28 degrees C. The plating efficiency was very high (67.54%) and doubling time was approximately 29 h. The kidney cell line was cryopreserved at different passage levels and revived successfully with 90-92% survival. Polymerase chain reaction amplification of mitochondrial 16S rRNA using primer specific to C striatus confirmed the origin of this cell line from murrel. The cell line was further characterized by chromosome number, transfection and mycoplasma detection. A marine fish nodavirus was tested to determine the susceptibility of this new cell line. The CSK cell line was found to be susceptible to nodavirus and the infection was confirmed by cytopathic effect (CPE), reverse transcriptase-polymerase chain reaction (RT-PCR), immunodot blot, enzyme linked immunosorbent assay (ELISA), virus replication efficiency and real time RT-PCR. The present study highlights the development and characterization of a new kidney cell line from an air breathing fish that could be used as an in vitro tools for propagation of fish viruses and gene expression studies. (C) 2013 Elsevier B.V. All rights reserved.