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Characterization of a novel brain cell line from Jian carp (Cyprinus carpio var. Jian)

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Abstract

Jian carp (Cyprinus carpio var. Jian) is an economically important cultured fish in China. Currently, it is facing threats from infectious diseases including koi herpesvirus (KHV). Here, we established a new cell line, designated CCB-J, derived from the brain tissue of the Jian carp. CCB-J cells grew well in Leibovitz’s L-15 medium containing 20% fetal bovine serum at 25 °C and have been subcultured for more than 60 passages. At the 30th passage, analysis showed that the number of chromosomes was 100, which is identical to that of other carp variants. Sequencing of the 18S ribosomal DNA confirmed that CCB-J originated from Jian carp. After transfection with the pEGFP-N1 plasmid, green fluorescence was observed in CCB-J. The replication of KHV in CCB-J cells was confirmed by RT-PCR and transmission electron microscopy. The viral titers of KHV in CCB-J cells and CCB cells, which have been widely used in the study of KHV, reached 103.9 and 101.8 median tissue culture infectious dose (TCID50/mL), respectively, within 14 days. The result of TaqMan PCR revealed that CCB-J cells were more sensitive to KHV than CCB cells. Meanwhile, a cytopathic effect (CPE) was also observed in the CCB-J cells in a shorter time post-infection compared with CCB cells. In summary, the CCB-J cell line will be a useful tool in the study of viral pathogenesis and vaccine research.

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Acknowledgments

We are grateful to Mr. Junming Cao, Guangdong Ocean University, China, for our support and encouragement.

Funding

This study was funded by the Science and Technology Planning Project of Guangdong Province, China (2016A020210029), Science and Technology Planning Project of Guangzhou City (201707010216), Guangdong Province Marine and Fishery Special Project (A201701C04), and National Natural Science Foundation of China (Grant No. 31402347).

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Authors and Affiliations

  1. College of Veterinary Medicine, South China Agricultural University, Guangzhou, 510642, China

    Ying Li & Yugu Li

  2. Institute of Animal Health, Guangdong Academy of Agricultural Sciences, Guangzhou, 510640, China

    Yanping Ma, Le Hao, Jiangyao Ma, Zhiling Liang, Zhenxing Liu & Hao Ke

  3. Key Laboratory of Livestock Disease Prevention of Guangdong Province, Guangzhou, 510640, China

    Yanping Ma, Le Hao, Jiangyao Ma, Zhiling Liang, Zhenxing Liu & Hao Ke

  4. Scientific Observation and Experiment Station of Veterinary Drugs and Diagnostic Techniques of Guangdong Province, Ministry of Agriculture and Rural Affairs, PRC, Guangzhou, 510640, China

    Yanping Ma, Le Hao, Jiangyao Ma, Zhiling Liang, Zhenxing Liu & Hao Ke

Authors
  1. Ying Li
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  2. Yanping Ma
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  3. Le Hao
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  4. Jiangyao Ma
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  5. Zhiling Liang
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  6. Zhenxing Liu
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  7. Hao Ke
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  8. Yugu Li
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Contributions

All authors contributed to the study conception and design. Ying Li developed the cell line and performed related experiments. Ying Li was also responsible for drafting the manuscript, as well as the acquisition, analysis, and interpretation of data. Yanping Ma participated in the verification and interpreted the data, and wrote parts of the manuscript. Le Hao analyzed the discussion and participated in the revision of the manuscript. Jiangyao Ma contributed to the conception and design of the current study. Zhiling Liang provided study materials and collected literature. Zhenxing Liu planned the main part of the experiments and assisted in the discussion of the manuscript. Zhenxing Liu was also the principal investigator of the funds. Hao Ke participated in the coordination of the study and reviewed the manuscript. Yugu Li plays a guiding role in this study, responsible for ensuring that the descriptions are accurate and agreed upon by all authors. All authors commented on previous versions of the manuscript. All authors read and approved the final manuscript.

Corresponding authors

Correspondence to Zhenxing Liu or Yugu Li.

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Li, Y., Ma, Y., Hao, L. et al. Characterization of a novel brain cell line from Jian carp (Cyprinus carpio var. Jian). Fish Physiol Biochem 47, 439–449 (2021). https://doi.org/10.1007/s10695-020-00923-4

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  • DOI: https://doi.org/10.1007/s10695-020-00923-4

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