Establishment, Characterization, and Virus Susceptibility of a New Marine Cell Line from Red Spotted Grouper (Epinephelus akaara)

Abstract

A marine fish cell line from the snout of red spotted grouper Epinephelus akaara, a protogynous hermaphrodite, was established, characterized, and subcultured with more than 60 passages. The grouper snout cell line (GSC) cells multiplied well in Dulbecco_s modified Eagle_s medium (DMEM) medium supplemented with 10% fetal bovine serum. The optimal growth temperature was 25-C, and morphologically the cells were fibroblastic. Chromosome analysis revealed that the GSC cell line has a normal diploid karyotype with 2n = 8st + 40t. A virus titration study indicated that the cells were susceptible to turbot Scophthalmus Maximus rhabdovirus (SMRV) (10 8.5 TCID 50 ml _ 1 ), while the viral titer of frog Rana grylio virus 9807 (RGV 9807 ) reached 10 3.5 TCID 50 ml _ 1 . The infection was confirmed by cytopathic effect (CPE), immunofluorescence, and electron microscopy experiments, which detected the viral particles in the cytoplasm of virus-infected cells, respectively. Further, significant fluorescent signals were observed when the GSC cells were transfected with pEGFP vector DNA, indicating their potential utility for transgenic and genetic manipulation studies.

Characterization of two grouper Epinephelus akaara cell lines: Application to studies of Singapore grouper iridovirus (SGIV) propagation and virus–host interaction

“…Grouper snout cell line (GSC) derived from grouper, E. akaara exhibited a chromosome number of 48 (Zhou et al, 2007). However, in our study, the chromosome number of EAGS and EAGSB cells was 88 and 72, respectively.…”

“…These two cell lines were well adapted to grow in L-15 medium supplemented with 10% FBS. L-15 medium was commonly used by different investigators for grouper cell cultures (Lai et al, 2003), while GSC cell lines derived from snout of grouper, E. akaara, was grown well in DMEM (Zhou et al, 2007). The suitable medium for EAGS and GAGSB cell cultures doesn't contain special requirements, such as NaCl addition, which is needed for other marine fish cells (Fernandez et al, 1993;Chew-Lim et al, 1994).…”

“…The establishment of healthy fish cell lines is essential for isolation, identification and characterization of infectious viruses from fishes (Parameswaran et al, 2007;Dong et al, 2008). Although a large number of cell lines (more than 160) have been established, relatively only a few cell lines were derived from marine fish (Fryer and Lannan, 1994;Chen et al, 2003;Bryson et al, 2006;Buonocore et al, 2006;Parameswaran et al, 2007;Zhou et al, 2007). Consequently, susceptible cell lines need to be established from marine fish for use in viral diagnostics.…”

Development of an Ussuri catfish Pseudobagrus ussuriensis skin cell line displaying differential cytopathic effects to three aquatic animal viruses

“…Development of diverse and sensitive fish cell lines is of priority interest for isolating and identifying fish viruses (Imajoh et al, 2007;Rougée et al, 2007;Zhou et al, 2007;Dong et al, 2011). Till now, more than 280 fish cell lines have been established, most of which are derived from model fishes or traditional cultured species, such as zebrafish (Brachydanio rerio), rainbow trout (Oncorhynchus mykiss), grass carp and groupers (Lakra et al, 2011).…”

RAPD-PCR analysis for molecular characterization and genotoxic studies of a new marine fish cell line derived from Dicentrarchus labrax

“…However, most of these cell lines are derived from freshwater fish species. The study on marine fish cell lines has developed rapidly in recent years and numerous cell lines from tissues of commercially important marine fish have been described (Buonocore et al 2005;Parameswaran et al 2007;Zhou et al 2007, Huang et al 2009Fan et al 2010). The advantages of these cell lines with respect to mammalian cells are that they are standardizable, easy to handle with relatively low variability, more convenient, and less laborious to use.…”