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Cytotechnology

, Volume 65, Issue 3, pp 307–322 | Cite as

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

  • P. Jayesh
  • Seena Jose
  • Rosamma Philip
  • I. S. Bright SinghEmail author
Method in Cell Science

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”.

Keywords

Shrimp cell line Penaeus monodon Shrimp cell culture medium (SCCM) MTT assay Lymphoid cell culture 

Notes

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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Copyright information

© Springer Science+Business Media B.V. 2012

Authors and Affiliations

  • P. Jayesh
    • 1
  • Seena Jose
    • 1
  • Rosamma Philip
    • 2
  • I. S. Bright Singh
    • 1
    Email author
  1. 1.National Centre for Aquatic Animal HealthCochin University of Science and TechnologyKochiIndia
  2. 2.Department of Marine Biology, Microbiology and Biochemistry, School of Marine SciencesCochin University of Science and TechnologyKochiIndia

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