Cell Proliferation – 3-(4,5-Dimethylthiazol-2-yl)-2,5-Diphenyltetrazolium Bromide (MTT) based assay
Activated HSCs are grown at about 70% confluency in 24 well plate. Next cells are subjected to drug treatment for 8 hr (standardization for individual drug). After incubation MTT is added to medium to get a final concentration of 1mg/ml. After 2 hr incubation with MTT,cells are subjected to DMSO treatment. DMSO lyse the cells to release MTT based colour products from the cells. The optical density (O.D.) of the colour products is measured by using the ELISA microplate reader at 570nm. (Majumder et al 2007 and Vistica et al 1991).
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2. Vistica DT, Skehan P, Scudiero D, Monks A, Pittman A, Boyd MR.Tetrazolium-based assays for cellular viability: a critical examination of selected parameters affecting formazan production.Cancer Res. 1991 May 15;51(10):2515-20.