Establishment and characterization of a continuous cell line from heart of Nile tilapia Oreochromis niloticus and its susceptibility to tilapia lake virus

Abstract

In the present study, we have developed a continuous cell line from the heart tissue of the Oreochromis niloticus and used for studying susceptibility to tilapia lake virus (TiLV). The cell line, designated as OnH, has been subcultured up to 82 passages. The optimal growth of OnH cells was observed at 28–32 °C in iL-15 medium supplemented with 20 % fetal bovine serum. Karyotype analysis revealed that the modal chromosome number of OnH cells was 44. Partial amplification and sequencing of 16S rRNA gene confirmed the origin of OnH cell line from O. niloticus. Immunophenotyping revealed that OnH cells were of epithelial origin. These cells were successfully transfected with pAcGFP1-N1 mammalian expression vector. OnH cells showed cytopathic effects following inoculation with TiLV. The virus titration study indicated that the cells were highly susceptible to TiLV with TCID50 value of 105.3/mL. The qRT-PCR studies revealed that the optimal temperature for TiLV replication in OnH cells was 28 °C. Further, transmission electron microscopy of TiLV-infected OnH cells showed a number of electron-dense virus particles measuring 60−90 nm diameter, which were enclosed in the vesicles in the cytoplasm. Therefore, the newly established OnH cell line provides a valuable tool for isolation of viruses from disease cases suspected to be of viral etiology in this candidate species' and also for transgenic and genetic manipulation studies.