Bright Singh, I S; Rosamma, Philip; Jayesh, P(Springer, February 25, 2014)
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Abstract:
Development of continuous cell lines from
shrimp is essential to investigate viral pathogens.
Unfortunately, there is no valid cell line developed
from crustaceans in general and shrimps in particular to
address this issue. Lack of information on the requirements
of cells in vitro limits the success of developing a
cell line, where the microenvironment of a cell culture,
provided by the growthmedium, is of prime importance.
Screening and optimization of growth medium components
based on statistical experimental designs have
been widely used for improving the efficacy of cell
culture media. Accordingly, we applied Plackett–Burman
design and response surface methodology to study
multifactorial interactions between the growth factors in
shrimp cell culture medium and to identify the most
important ones for growth of lymphoid cell culture from
Penaeus monodon. The statistical screening and
optimization indicated that insulin like growth factor-I
(IGF-I) and insulin like growth factor-II (IGF-II) at
concentrations of 100 and 150 ng ml-1, respectively,
could significantly influence the metabolic activity and
DNA synthesis of the lymphoid cells. An increase of
53 % metabolic activity and 24.8 % DNA synthesis
could be obtained, which suggested that IGF-I and IGFII
had critical roles in metabolic activity and DNA
synthesis of shrimp lymphoid cells
Bright Singh, I S; Rosamma, Philip; Jayesh, P; Seena, Jose(Springer, September 30, 2012)
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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
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’’.
Description:
Cytotechnology (2013) 65:307–322
DOI 10.1007/s10616-012-9491-9