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Cloning of a new fibroblast cell line from an early primary culture from mandarin fish (Siniperca chuatsi) fry for efficient proliferation of megalocytiviruses

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Abstract

Megalocytiviruses are important emerging pathogens in both freshwater and marine finfish aquaculture. However, a limited number of piscine cell lines are persistently susceptible to these viruses, which greatly limits the study of megalocytiviruses. In this study, a new fibroblast-like cell line was established from an early primary culture from mandarin fish fry by a single cell cloning and was designated as MFF-8C1. The MFF-8C1 cells grow well in Dulbecco’s modified Eagle’s medium supplemented with 10 % fetal bovine serum and had been subcultured more than 60 passages since the initial recovery culture in October 2009. Chromosomal analysis revealed that 91 % of the MFF-8C1 cells maintained a normal diploid chromosome number (2n = 48) in the 46th passage. Infection experiments showed that both freshwater-borne and marine-borne megalocytiviruses induce severe cytopathic effects in infected MFF-8C1 cells characterized by the rounding and enlargement of cells, which are highly consistent with the previous description of the infection in other susceptible cells with megalocytivirus. Megalocytivirus infections were further confirmed by a transmission electron microscopy. Furthermore, the MFF-8C1-cultured megalocytiviral suspension was highly virulent to infected mandarin fish. In summary, a new fibroblast cell line from mandarin fish fry that was highly permissive to megalocytiviruses was established. The MFF-8C1 cell line is a promising cellular substrate candidate for cell-cultured vaccine production of megalocytivirus.

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Acknowledgments

This research was supported by the National Basic Research Program of China under Grant No. 2012CB114406; Technology Planning Project of Guangdong Province under number 2011A020102002 and 2012A020800006, and the planning subject of ‘the twelfth five-year-plan’ in national science and technology for the rural development in China under number 2011BAD13B00.

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Authors and Affiliations

  1. MOE Key Laboratory of Aquatic Food Safety/State Key Laboratory for Bio-control, School of Life Sciences, Sun Yat-sen (Zhongshan) University, Guangzhou, 510275, People’s Republic of China

    Chuanfu Dong, Fan Shuang, Shaoping Weng & Jianguo He

  2. School of Marine Sciences, Sun Yat-sen University, Guangzhou, 510275, People’s Republic of China

    Jianguo He

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  1. Chuanfu Dong
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  2. Fan Shuang
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  3. Shaoping Weng
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  4. Jianguo He
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Correspondence to Chuanfu Dong.

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Dong, C., Shuang, F., Weng, S. et al. Cloning of a new fibroblast cell line from an early primary culture from mandarin fish (Siniperca chuatsi) fry for efficient proliferation of megalocytiviruses. Cytotechnology 66, 883–890 (2014). https://doi.org/10.1007/s10616-013-9642-7

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  • DOI: https://doi.org/10.1007/s10616-013-9642-7

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