title: Establishment and characterization of a new cell line derived from kidney of grouper, Epinephelus akaara (Temminck & Schlegel), susceptible to Singapore grouper iridovirus (SGIV).
abstract: A marine fish cell line derived from the kidney of red-spotted grouper, Epinephelus akaara, designated as EAGK was established and characterized. The EAGK cells multiplied well in Leibovitz's L-15 medium containing 10% foetal bovine serum at 25 °C and have been subcultured for more than 90 passages. Karyotyping, chromosomal typing and ribosomal RNA (rRNA) genotyping analysis revealed that EAGK had a modal diploid chromosome number of 82 and was a fibroblast cell line originated from grouper. A severe cytopathic effect was observed in EAGK cells incubated with Singapore grouper iridovirus (SGIV), but not with soft-shelled turtle iridovirus, viral nervous necrosis virus or spring viraemia of carp virus. SGIV replication was further confirmed by immunofluorescence, electron microscopy and virus titre determination. Bright fluorescence was observed after transfection with fluorescent protein reporter plasmids, indicating that EAGK cells can be used to identify gene functions in vitro. In addition, the cell organelles including mitochondria and endoplasm reticulum changed and aggregated around virus factories after SGIV infection, suggested that the EAGK cell line could be an important tool for investigation of iridovirus-host interactions.
authors.affiliation: Key Laboratory of Marine Bio-Resources Sustainable Utilization, South China Sea Institute of Oceanology, The Chinese Academy of Sciences, Guangzhou, China.
authors.forename: J
authors.initials: J
authors.lastname: Gong
authors.forename: Y
authors.initials: Y
authors.lastname: Huang
authors.forename: X
authors.initials: X
authors.lastname: Huang
authors.forename: Z
authors.initials: Z
authors.lastname: Ouyang
authors.forename: M
authors.initials: M
authors.lastname: Guo
authors.forename: Q
authors.initials: Q
authors.lastname: Qin
mesh_terms: D000818:Animals; D001483:Base Sequence; D002460:Cell Line; D002875:Chromosomes / Q000235:genetics*; D015925:Cryopreservation; D017931:DNA Primers / Q000235:genetics; D007621:Karyotyping; D007668:Kidney / Q000166:cytology*; D008854:Microscopy, Electron; D008856:Microscopy, Fluorescence; D008969:Molecular Sequence Data; D010473:Perciformes / Q000235:genetics / Q000821:virology*; D061251:Primary Cell Culture / Q000379:methods*; D012335:RNA, Ribosomal / Q000235:genetics; D017995:Ranavirus; D017422:Sequence Analysis, DNA
chemical_list: D017931:DNA Primers; D012335:RNA, Ribosomal