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Abstract:
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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
8C. 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 (29), 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’’. |