Establishment of a cell line from swim bladder of the Grass carp (Ctenopharyngodon idellus) for propagation of Grass Carp Reovirus Genotype II.

Affiliation

Yang Y(1), Wang Y(2), Wang Q(3), Zeng W(4), Li Y(5), Yin J(6), Wu S(7), Shi C(8).
Author information:
(1)Key Laboratory of Fishery Drug Development of Ministry of Agriculture, Key Laboratory of Aquatic Animal Immune Technology of Guangdong Province, Pearl River Fisheries Research Institute, Chinese Academy of Fishery Sciences, Guangzhou, Guangdong, PR China; College of Fisheries, Tianjin Agricultural University, Tianjin, 300384, China. Electronic address: [Email]
(2)Key Laboratory of Fishery Drug Development of Ministry of Agriculture, Key Laboratory of Aquatic Animal Immune Technology of Guangdong Province, Pearl River Fisheries Research Institute, Chinese Academy of Fishery Sciences, Guangzhou, Guangdong, PR China. Electronic address: [Email]
(3)Key Laboratory of Fishery Drug Development of Ministry of Agriculture, Key Laboratory of Aquatic Animal Immune Technology of Guangdong Province, Pearl River Fisheries Research Institute, Chinese Academy of Fishery Sciences, Guangzhou, Guangdong, PR China. Electronic address: [Email]
(4)Key Laboratory of Fishery Drug Development of Ministry of Agriculture, Key Laboratory of Aquatic Animal Immune Technology of Guangdong Province, Pearl River Fisheries Research Institute, Chinese Academy of Fishery Sciences, Guangzhou, Guangdong, PR China; Guangdong Provincial Key Laboratory of Animal Molecular Design and Precise Breeding, Key Laboratory of Animal Molecular Design and Precise Breeding of Guangdong Higher Education Institutes, School of Life Science and Engineering, Foshan University, Foshan, 440605, China. Electronic address: [Email]
(5)Key Laboratory of Fishery Drug Development of Ministry of Agriculture, Key Laboratory of Aquatic Animal Immune Technology of Guangdong Province, Pearl River Fisheries Research Institute, Chinese Academy of Fishery Sciences, Guangzhou, Guangdong, PR China. Electronic address: [Email]
(6)Key Laboratory of Fishery Drug Development of Ministry of Agriculture, Key Laboratory of Aquatic Animal Immune Technology of Guangdong Province, Pearl River Fisheries Research Institute, Chinese Academy of Fishery Sciences, Guangzhou, Guangdong, PR China. Electronic address: [Email]
(7)Key Laboratory of Fishery Drug Development of Ministry of Agriculture, Key Laboratory of Aquatic Animal Immune Technology of Guangdong Province, Pearl River Fisheries Research Institute, Chinese Academy of Fishery Sciences, Guangzhou, Guangdong, PR China. Electronic address: [Email]
(8)Key Laboratory of Fishery Drug Development of Ministry of Agriculture, Key Laboratory of Aquatic Animal Immune Technology of Guangdong Province, Pearl River Fisheries Research Institute, Chinese Academy of Fishery Sciences, Guangzhou, Guangdong, PR China. Electronic address: [Email]

Abstract

A cell line was established from swim bladder of the Grass carp (Ctenopharyngodon idellus) (CiSB), which was permissive for infection and propagation of Grass Carp Reovirus (GCRV). CiSB cells displayed optimal growth at 27 °C using M199 medium containing 10% fetal bovine serum and a fibroblastic-like morphology. Karyotype analysis revealed that the average diploid chromosome number was 52 in 58% of cells at passage 60 compared to the wild type Grass carp cells (2n = 48). Infection with GCRV II isolate Hunan1307 was tracked by immunofluorescence and virus titration assay. The virus titer reached 105.2 TCID50/mL on 7th days post infection (dpi). Healthy adult Grass carp that were challenged with the virus propagated onto CiSB cells, displayed the typical symptoms and histopathological changes of Grass carp hemorrhagic disease (GCHD). Therefore, the CiSB cells can be used to propagate GCRV II and serve as a useful tool to study the pathogenesis of GCHD.