2.4 Determination of In vitro cytotoxic effect on
cultured Cells
L929/A549 cell lines were purchased from National Centre for Cell
Science (NCCS), Pune, India. Cultures were maintained in Dulbecco’s
modified eagles media (Himedia, India) supplemented with 10% fetal
bovine serum (FBS) (Invitrogen) and grown to confluence at 37 °C and 5%
CO2 in a humidified atmosphere using a
CO2 incubator (NBS, Eppendorf, Germany). The cells were
trypsinised (500 µL of 0.025% Trypsin in PBS/0.5 mM EDTA solution
(Himedia, India) for 2 mts and passed to T flasks under aseptic
condition. Test compounds were added to grown cells at a final
concentration of 6.25, 12.5, 25, 50 and 100 µg/mL from a stock of 1
mg/mL and incubated for 24 hrs. The percentage difference in viability
was determined by standard MTT assay (Arung et al ., 2000) after
24 hrs of incubation.