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Establishment and Characterization of a Novel Kidney-cell Line from Orange-spotted Grouper, Epinephelus coioides, and its Susceptibility to Grouper Iridovirus | Abstract
ISSN: 2155-9910

Journal of Marine Science: Research & Development
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Research Article

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

Sue-Min Huang1,2, Chien Tu1, Shu-Ting Kuo1, Hung-Chih Kuo3, Chi-Chung Chou4 and Shao-Kuang Chang2*

1Animal Health Research Institute, Council of Agriculture, Taipei, Taiwan

2Institute of Veterinary Medicine, School of Veterinary Medicine, National Taiwan University, Taipei, Taiwan

3Department of Veterinary Medicine, National Chiayi University, Chiayi, Taiwan

4Department of Veterinary Medicine, National Chung-Hsing University, Taichung, Taiwan

*Corresponding Author:
Chang SK
Institute of Veterinary Medicine, School of Veterinary Medicine
National Taiwan University, Taipei 10617, Taiwan
Tel: +886- 2-3366-3864
E-mail: [email protected]

Received date: April 07, 2016; Accepted date: May 23, 2016; Published date: May 31, 2016

Citation: Huang SM, Tu C, Kuo ST, Kuo HC, Chou CC, et al. (2016) Establishment and Characterization of a Novel Kidney-cell Line from Orange-spotted Grouper, Epinephelus coioides, and its Susceptibility to Grouper Iridovirus. J Marine Sci Res Dev 6:192. doi:10.4172/2155-9910.1000192

Copyright: © 2016 Huang SM, et al. This is an open-access article distributed under the terms of the Creative Commons Attribution License, which permits unrestricted use, distribution, and reproduction in any medium, provided the original author and source are credited.

Abstract

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 108.4 TCID50 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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Citations : 1719

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