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A novel medium for the development of in vitro cell culture system from Penaeus monodon

  • Method in Cell Science
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Abstract

Lack of a valid shrimp cell line has been hampering the progress of research on shrimp viruses. One of the reasons identified was the absence of an appropriate medium which would satisfy the requirements of the cells in vitro. We report the first attempt to formulate an exclusive shrimp cell culture medium (SCCM) based on the haemolymph components of Penaeus monodon prepared in isosmotic seawater having 27 ‰ salinity. The SCCM is composed of 22 amino acids, 4 sugars, 6 vitamins, cholesterol, FBS, phenol red, three antibiotics, potassium dihydrogen phosphate and di-sodium hydrogen phosphate at pH 6.8–7.2. Osmolality was adjusted to 720 ± 10 mOsm kg−1 and temperature of incubation was 25 ºC. The most appropriate composition was finally selected based on the extent of attachment of cells and their proliferation by visual observation. Metabolic activity of cultured cells was measured by MTT assay and compared with that in L-15 (2×), modified L-15 and Grace’s insect medium, and found better performance in SCCM especially for lymphoid cells with 107 % increase in activity and 85 ± 9 days of longevity. The cells from ovary and lymphoid organs were passaged twice using the newly designed shrimp cell dissociation “cocktail”.

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Acknowledgments

This research was supported by Department of Biotechnology, Government of India (BT/PR8050/AAQ/03/289/2006). The first author thanks DBT for Fellowship.

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Authors and Affiliations

  1. National Centre for Aquatic Animal Health, Cochin University of Science and Technology, Fine Arts Avenue, Kochi, 682016, India

    P. Jayesh, Seena Jose & I. S. Bright Singh

  2. Department of Marine Biology, Microbiology and Biochemistry, School of Marine Sciences, Cochin University of Science and Technology, Fine Arts Avenue, Kochi, 682016, India

    Rosamma Philip

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  1. P. Jayesh
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  2. Seena Jose
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  4. I. S. Bright Singh
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Correspondence to I. S. Bright Singh.

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Jayesh, P., Jose, S., Philip, R. et al. A novel medium for the development of in vitro cell culture system from Penaeus monodon . Cytotechnology 65, 307–322 (2013). https://doi.org/10.1007/s10616-012-9491-9

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