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Establishment and characterization of a new marine fish cell line derived from red-spotted grouper Epinephelus akaara

Authors

  • Z. L. Ou-yang,

    1. State Key Laboratory for Biocontrol, School of Life Sciences, Sun Yat-sen University, 135 West Xingang Road, Guangzhou 510275, P.R. China
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  • X. H. Huang,

    1. Key Laboratory of Marine Bio-resources Sustainable Utilization, South China Sea Institute of Oceanology, Chinese Academy of Sciences, 164 West Xingang Road, Guangzhou 510301, P.R. China
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  • E. Y. Huang,

    1. State Key Laboratory for Biocontrol, School of Life Sciences, Sun Yat-sen University, 135 West Xingang Road, Guangzhou 510275, P.R. China
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  • Y. H. Huang,

    1. Key Laboratory of Marine Bio-resources Sustainable Utilization, South China Sea Institute of Oceanology, Chinese Academy of Sciences, 164 West Xingang Road, Guangzhou 510301, P.R. China
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  • J. Gong,

    1. State Key Laboratory for Biocontrol, School of Life Sciences, Sun Yat-sen University, 135 West Xingang Road, Guangzhou 510275, P.R. China
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  • J. J. Sun,

    1. State Key Laboratory for Biocontrol, School of Life Sciences, Sun Yat-sen University, 135 West Xingang Road, Guangzhou 510275, P.R. China
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  • Q. W. Qin

    Corresponding author
    1. Key Laboratory of Marine Bio-resources Sustainable Utilization, South China Sea Institute of Oceanology, Chinese Academy of Sciences, 164 West Xingang Road, Guangzhou 510301, P.R. China
      Tel.–fax: +86 20 89023638; email: qinqw@scsio.ac.cn
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Tel.–fax: +86 20 89023638; email: qinqw@scsio.ac.cn

Abstract

A new marine fish cell line, EAGL, derived from the liver of red-spotted grouper Epinephelus akaara was established and characterized. The cells multiplied well in minimum essential medium (MEM) supplemented with 10% foetal bovine serum (FBS) at temperatures between 25 and 30° C. The growth rate of this cell line increased as the proportion of FBS increased from 5 to 20% at 25° C, with maximum growth at the concentration of 15 or 20% FBS. Morphologically, the cells were epithelial-like and the presence of pancytokeratin confirmed their epithelial origin. Chromosome analysis revealed that the modal chromosome number was 48. The susceptibility of the cell line to four fish viruses was tested. Significant cytopathic effect (CPE) was only observed in Singapore grouper iridovirus (SGIV)-infected cells, and the virus replication was further confirmed by immunofluorescence, electron microscopy and real-time reverse-transcription (RT)-PCR assay. When the cells were transfected with pEGFP-N3 plasmid, bright fluorescent signals were observed, suggesting that this cell line can be used for transgenic and genetic manipulation studies.

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