Establishment of an astrocyte-like cell line from the brain of tilapia (Oreochromis niloticus) for virus pathogenesis and a vitro model of the blood-brain barrier

J Fish Dis. 2022 Oct;45(10):1451-1462. doi: 10.1111/jfd.13674. Epub 2022 Jun 27.

Abstract

A new cell line was established from the brain of a cultured fish, tilapia (Oreochromis niloticus), designated as TA-02 (Tilapia Astrocyte clone 02 cell line). The TA-02 cells are grown for 300 days in an L-15 medium supplemented with 10% fetal bovine serum (FBS). This cell line showed excellent proliferative capacity and expressed various neuroglial cell markers, including SOX2, SOX10, Hes1, Notch1, Occludin, E-cadherin, and GFAP. In addition, TA-02 cells were susceptible to Tilapia Lake Virus (TiLV) as demonstrated by the presence of a severe cytopathic effect (CPE), virus particle in a transmission electron microscope (TEM), and PCR positive signal. Bacterial cytotoxicity studies showed that Streptococcus agalactiae was toxic to TA-02 cells. When co-culture with trans-well, TA-02 exhibited prominent barrier properties, manifested by tight intercellular junctions and increased trans-endothelial electrical resistance (TEER). In addition, the barrier is effective against Escherichia coli (non-meningitis pathogenic bacteria). In contrast, S. agalactiae (meningitis pathogenic bacteria) can pass through the membrane comprising the cells in the trans-well insert. The newly established TA-02 cell line provided a valuable tool for virus pathogenesis and a vitro model of the fish blood-brain barrier.

Keywords: Oreochromis niloticus; astrocyte; blood-brain barrier; cell line; trans-well.

MeSH terms

  • Animals
  • Astrocytes
  • Bacteria
  • Blood-Brain Barrier
  • Brain
  • Cell Line
  • Cichlids* / microbiology
  • Fish Diseases* / microbiology
  • Streptococcal Infections* / veterinary
  • Streptococcus agalactiae
  • Tilapia*