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Development and Characterization of Two New Cell Lines from Milkfish (Chanos chanos) and Grouper (Epinephelus coioides) for Virus Isolation

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Abstract

Two new cell lines, SIMH and SIGE, were derived from the heart of milkfish (Chanos chanos), a euryhaline teleost, and from the eye of grouper (Epinephelus coioides), respectively. These cell lines were maintained in Leibovitz’s L-15 supplemented with 20% fetal bovine serum (FBS). The SIMH cell line was subcultured more than 50 times over a period of 210 days and SIGE cell line has been subcultured 100 times over a period of 1 1/2 years. The SIMH cell line consists predominantly of fibroblastic-like cells. The SIGE cell line consists predominantly of epithelial cells. Both the cell lines were able to grow at temperatures between 25 and 32°C with an optimum temperature of 28°C. The growth rate of these cells increased as the proportion of FBS increased from 2% to 20% at 28°C with optimum growth at the concentrations of 15% or 20% FBS. Seven marine fish viruses were tested to determine the susceptibility of these cell lines. The SIGE cell line was found to be susceptible to nodavirus, MABV NC-1 and Y6, and the infection was confirmed by cytopathic effect (CPE) and reverse transcriptase-polymerase chain reaction. When these cells were transfected with pEGFP-N1 vector DNA, significant fluorescent signals were observed, suggesting that these cell lines can be a useful tool for transgenic and genetic manipulation studies. Further, these cell lines are characterized by immunocytochemistry using confocal laser scanning microscopy (CFLSM).

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Acknowledgment

The authors thank the administrators of C. Abdul Hakeem College for providing the facilities to carry out this work. The authors also thank the Director, Central Institute of Brackishwater Aquaculture for providing the experimental animals. This work was funded by Department of Biotechnology, Government of India, New Delhi, India.

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Authors and Affiliations

  1. Aquaculture Biotechnology Division, Department of Zoology, C.Abdul Hakeem College, Melvisharam-632 509, Vellore Dt., Tamilnadu, India

    V. Parameswaran, V. P. Ishaq Ahmed & A. S. Sahul Hameed

  2. National Centre for Cell Science, Pune University Campus, Ganeshkhind, Pune, India

    Ravi Shukla & R. R. Bhonde

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  1. V. Parameswaran
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  2. V. P. Ishaq Ahmed
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  3. Ravi Shukla
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  4. R. R. Bhonde
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  5. A. S. Sahul Hameed
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Correspondence to A. S. Sahul Hameed.

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Parameswaran, V., Ishaq Ahmed, V.P., Shukla, R. et al. Development and Characterization of Two New Cell Lines from Milkfish (Chanos chanos) and Grouper (Epinephelus coioides) for Virus Isolation. Mar Biotechnol 9, 281–291 (2007). https://doi.org/10.1007/s10126-006-6110-9

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