Establishment and characterization of a continuous marine fish fin cell line from <italic style="font-style: italic">Epinephelus</italic> <italic style="font-style: italic">fuscoguttatus</italic> and viral susceptibility to crustacean CMNV

Ting XUE; Yue ZHANG; Zhenxin ZHAO; Yaru LI; Jinwu WANG; Huarong GUO

doi:10.1007/s00343-024-4054-4

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Establishment and characterization of a continuous marine fish fin cell line from Epinephelus fuscoguttatus and viral susceptibility to crustacean CMNV

Biology | Updated：2025-05-27

- Establishment and characterization of a continuous marine fish fin cell line from Epinephelus fuscoguttatus and viral susceptibility to crustacean CMNV
- Journal of Oceanology and Limnology Vol. 43, Issue 3, Pages: 892-905(2025)
- Affiliations：
  
  1.Key Laboratory of Marine Genetics and Breeding (Ministry of Education) and College of Marine Life Sciences, Ocean University of China, Qingdao 266003, China
  2.Key Laboratory of Evolution & Marine Biodiversity (Ministry of Education) and Institute of Evolution & Marine Biodiversity, Ocean University of China, Qingdao 266003, China
- Author bio：
  
  huarongguo@ouc.edu.cn
- Funds：
- DOI：10.1007/s00343-024-4054-4
  CLC：
- Received：29 February 2024，
  
  Online First：20 May 2024，
  
  Published：01 May 2025
- Accepted：
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XUE Ting,ZHANG Yue,ZHAO Zhenxin,et al.Establishment and characterization of a continuous marine fish fin cell line from Epinephelus fuscoguttatus and viral susceptibility to crustacean CMNV[J].Journal of Oceanology and Limnology,2025,43(03):892-905.
DOI：

XUE Ting,ZHANG Yue,ZHAO Zhenxin,et al.Establishment and characterization of a continuous marine fish fin cell line from Epinephelus fuscoguttatus and viral susceptibility to crustacean CMNV[J].Journal of Oceanology and Limnology,2025,43(03):892-905. DOI： 10.1007/s00343-024-4054-4.

摘要

Abstract

Fish cell line provide a useful tool for studies in virology and molecular biology. To establish a novel continuous marine fish cell line (EFF) from caudal fin tissue of brown-marbled grouper

Epinephelus

fuscoguttatus

its susceptibility to the crustacean covert mortality nodavirus (CMNV) was evaluated. The primary cell cultures were initiated first by incomplete digestion of fin tissue blocks with dispase and collagenase

and then the explant was cultured in L-15 medium supplemented with 20% fetal bovine serum

10% grouper muscle extract

and 20-ng/mL growth factors of basic fibroblast growth factor (bFGF) and epidermal growth factor (EGF). The EFF cells were continuously passaged beyond 50 times in fibroblast-like morphology

and they grew well in L-15 medium supplemented with a lower concentration fetal bovine serum (10%) at 28 °‍‍C after passage 10

without muscle extract and the growth factors. In addition

their grouper origin was confirmed by chromosome analysis and cytochrome oxidase 1 (

CO1

) gene analysis of these EFF cells. Transfection experiment via lipofectamine 8 000 indicated that the EFF cells had a high transfection potential with a transfection efficiency up to 32%. Cross-species viral susceptibility analysis showed that

CMNV not only successfully infected the EFF cells as evidenced by obvious cytopathic effects like vacuolation

detachment and death of cells

but also multiplied in the EFF cells as indicated by the results of semi-quantitative RT-PCR in transmission electron microscopy. Therefore

the establishment of immortal EFF cell line provided a useful cell model for future works on the isolation

multiplication

and pathogenic mechanism of cross-species infection of CMNV as well as genetic manipulation.

关键词

Keywords

references

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Establishment and characterization of a continuous marine fish fin cell line from Epinephelus fuscoguttatus and viral susceptibility to crustacean CMNV

DOI：10.1007/s00343-024-4054-4

摘要

Abstract

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Keywords

references