A New Conditionally Immortalized Nile Tilapia (Oreochromis niloticus) Heart Cell Line: Establishment and Functional Characterization as a Promising Tool for Tilapia Myocarditis Studies

Abstract

A new cell line named the tilapia heart cell line (TAH-11) was established from the heart of tilapia (Oreochromis niloticus) by enzymatic digestion and mechanical separation. The TAH-11 cell line has been stably subcultured for over 80 generations and resuscitated after being frozen in liquid nitrogen for six months, with exuberant cell growth. The optimal culture condition of TAH-11 is L-15 medium supplemented with 10% fetal bovine serum at 25 °C. Sequencing of the mitochondrial 18S rRNAs confirmed that TAH-11 cells were derived from the tilapia. TAH-11 was also identified as a myocardial cell line based on the mRNA expression of the troponin I, α-actin and myoglobin genes. Simultaneously, TAH-11 could be successfully transfected with the GFP reporter gene, suggesting that the TAH-11 cell line could be used for exogenous gene expression in vitro. The TAH-11 cells showed susceptibility to CGSIV, which was demonstrated by the presence of a severe cytopathic effect, suggesting that the TAH-11 cell line is an ideal tool for studying host–virus interaction and potential vaccines. In addition, the expression of inflammatory factors in TAH-11 cells can be remarkably induced following Streptococcus agalactiae or Streptococcus iniae. The present data lays a foundation to further explore the mechanism of how Streptococcus spp. causes tilapia myocarditis.