Characterization and virus susceptibility of a skin cell line from red-spotted grouper (Epinephelus akaara)

Abstract

A red-spotted grouper Epinephelus akaara skin (RGS) cell line was established and characterized. RGS cells had a normal diploid chromosome number of 2n = 48, the morphology of which was fibroblastic-like in 3 days and epithelial-like over 5 after 16 passages. The cells multiplied well in Dulbecco's modified Eagle's medium supplemented with 10% of fetal bovine serum at 25°C. Susceptibilities of RGS and grass carp ovary (GCO) cells to two viruses were tested, and the results showed that the titer of an iridovirus Rana grylio virus (RGV) in RGS cells was 10³·⁵ TCID₅₀ ml⁻¹, which was much higher than a rhabdovirus spring viremia of carp virus (SVCV) in the cells (10⁰·⁵ TCID₅₀ ml⁻¹). The titers of RGV and SVCV in GCO were 10⁶·⁰ TCID₅₀ ml⁻¹ and 10⁸·⁰ TCID₅₀ ml⁻¹, respectively, which were higher than those in RGS cells. The data may imply that RGS cells could be selectively resistible to some viruses during infection. RT-PCR analysis of RGV-infected RGS cells showed that RGV could replicate in RGS cells. Further study of virus replications in RGS cells was conducted by electron microscopy and immunofluorescence microscopy has shown that virus particles scattered in the cytoplasm and virus protein appeared in both the cytoplasm and nucleus. The results suggested that RGS cells could be used as a potential in vitro model to study the cutaneous barrier function against virus infection.