Screening of Selected Chemotherapeutant by using Primary Liver cell Culture of Labeo rohita (Hamilton, 1822)
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Keywords:
Labeo rohita, primary cell culture, in vitro screening, InsecticideAbstract
Development of primary cell culture from liver
tissues of Labeo rohita was successfully completed by
following explant culture method. The L-15 media
supplemented with 20% foetal bovine serum (FBS)
at 28oC has resulted in good attachment and
proliferation of primary culture. FBS is required for
efficient attachment of liver explant but increased
supplementation of FBS in media did not stimulate
the proliferation of cells. Radiation from liver
explant was first seen after 48 h of attached explant.
The monolayer formation of liver cell culture was
observed after twenty one days and it was subsequently
sub cultured for more than 10 times. The in
vitro cytotoxicity of a commonly used drugs (Oxytetracycline),
chemotherapeutant (Benzalkonium chloride)
and insecticide (Fipronil) was studied on
primary liver cell culture of L. rohita by assessing cell
viability using tetrazolium (MTT) assay which
revealed that Fipronil is most toxic compared to OTC
and BKC. Reduction in cell viability was observed
with the increase in concentration of
chemotherapeutants. The IC50 values of OTC, BKC
and Fipronil calculated from dose-response curve
were 76.43 μg ml-1, 3.22 μg ml-1 and 0.014 μg ml-1
respectively. The study revealed that primary cell
culture from liver tissue of L. rohita can be used as
a potentially bio-indicator for the in vitro screening
of the acute toxicities of Fipronil.