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

Huang, Xiaohong; Huang, Youhua; Sun, Jingjing; Han, Xin; Qin, Qiwei

doi:10.1016/j.aquaculture.2009.04.019

Cited by 180 publications

(95 citation statements)

References 28 publications

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“…Some fish cell lines had been reported that a mixture of fibroblastic-like and epithelial-like cells were observed in initial passages (Lai et al 2003;Lakra et al 2006;Huang et al 2009;Ku et al 2010), which was similar with the RGS cells. However, the morphology of RGS cells changed during subculture, which has not been reported elsewhere.…”

Section: Discussionmentioning

confidence: 65%

“…Karyotype analysis revealed that 59% of the cells possessed a diploid chromosome number of 2n = 48, which was identical with the modal number of grouper cell lines such as GS, GSC, SIGE, and RGB (Qin et al 2006;Parameswaran et al 2007;Zhou et al 2007;Ku et al 2009), whereas some other cell lines derived from groupers, such as GSB, GH, GF, GBC1, EAGS, and EAGSB cells, and the chromosome numbers of them were 86, 66, 58, 44, 88, and 72, respectively (Lai et al 2003;Wen et al 2008;Huang et al 2009), which demonstrated that cell lines derived from different tissues of the same species may have varied number of chromosomes. These results may indicate the occurrence of aneuploidy due to cell immortalization.…”

Section: Discussionmentioning

confidence: 91%

“…Most of the established cell lines are derived from splanchnic organs, such as kidney, liver, brain, heart, spleen, embryo, and swim bladder (Bryson et al 2006;Ku et al 2009;Huang et al 2009;Ou-Yang et al 2010). However, cell lines derived from healthy skin of fish were rare.…”

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confidence: 99%

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

Chen

et al. 2012

Fish Physiol Biochem

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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 10 3.5 TCID 50 ml -1 , which was much higher than a rhabdovirus spring viremia of carp virus (SVCV) in the cells (10 0.5 TCID 50 ml -1 ). The titers of RGV and SVCV in GCO were 10 6.0 TCID 50 ml -1 and 10 8.0 TCID 50 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.

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Section: Discussionmentioning

confidence: 65%

Section: Discussionmentioning

confidence: 91%

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

Chen

et al. 2012

Fish Physiol Biochem

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“…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.…”

Section: Introductionmentioning

confidence: 99%

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

et al. 2013

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Continuous cell lines could provide an important tool for studying epidemiology, toxicology, cellular physiology and the host-pathogen interactions. Random amplified polymorphic deoxyribonucleic acid analysis by PCR (RAPD-PCR) was used for the molecular characterization of Dicentrarchus labrax embryonic cells (DLEC) as a possible tool to detect DNA alterations in environmental genotoxic studies. We studied the DNA pattern of the DLEC fish cell line, a fibroblast-like cell line derived from European sea bass. From a total of 15 primers only six showed good discriminatory power for the amplification process on DNA samples collected from cells by three different methods (organic extraction, saltingout method and chelating agent extraction). The results obtained show that the cell line chosen for this study could be used as a possible tool for the detection of potential genotoxicity of numerous chemical compounds.

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“…Hence, development a continuous cell line from tropical fish species for identifying pathogenesis of viral diseases and for vaccine production is imperative. To date, more than 22 cell lines have been developed from grouper for iridovirus and NNV isolation, such as seven cell lines established from eye, fin, heart, swimbladder, kidney, liver and brain of E. awoara; four from brain, fin, eye and spleen of E. coioides [3,[8][9][10][11][12][13][14][15][16][17][18][19][20][21][22][23]; five from snout, spleen, live, kidney and swimbladder of E. akaara [4,9,15,24,25]; three from brain, gill, heart of E. quoyanus [10]; one from heart of E. lanceolatus [5]; one from embryo cells of E. tauvina [17]; and one from fin of E. fuscoguttatus [22]. Although four cell lines derived from E. coioides, have been documented, no cell line has been derived from kidney of the species.…”

Section: Introductionmentioning

confidence: 99%

Establishment and Characterization of a Novel Kidney-cell Line from Orange-spotted Grouper, Epinephelus coioides, and its Susceptibility to Grouper Iridovirus

Tu¹,

Kuo²

2016

J Marine Sci Res Dev

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A new continuous cell line, designated as GK-7, was developed from the kidney tissue of the marine grouper, Epinephelus coioides. The cell line grew well in Leibovitz's L-15 medium supplemented with 15% fetal bovine serum at a range of temperatures from 20 to 32℃, with optimal growth at 25℃. Morphologically, the GK-7 cell line is spindleshaped epithelial-like cell comfirmed by immunophenotyping with cytokeratin antibody. Chromosome number analysis showed that GK-7 cells of the 50th and 150th cell passages had a modal diploid chromosome number of 48 and 66, respectively. Replication of GIV with the cell line showed that the maximum virus yield reached up 10 8.4TCID 50 mL -1 with the cells of the 50th passage. Electron micrographs showed abundant cytoplasmic icosahedral virions with a mean diameter of 200 nm in virus-infected cells. Negative staining of ultrathin sections of infected cells showed three-layered membrane enveloped mature viral particles with a diameter about 240 nm. Green fluorescent protein can be expressed in both cell lines at 48 hr after the cell lines were transfected with a green fluorescent reporter gene driven by a cytomegalovirus promoter. Our results showed that the GK-7 cell line provided valuable tools for the isolation and investigation of fish iridovirus and for vaccine production.

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

Cited by 180 publications

References 28 publications

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

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

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

Establishment and Characterization of a Novel Kidney-cell Line from Orange-spotted Grouper, Epinephelus coioides, and its Susceptibility to Grouper Iridovirus

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