Assessment of Anticancer Activity by MTT Assay

General anti-cancer agents, likewise chemotherapeutic medications or Taxol can suppress cell proliferation; therefore, the viability assay of these agents is expected to show almost the same results on any high proliferating cell line despite the cancerous or normal source of the cells. Consequently, researchers only determine the effects of such remedies on the specific tissue cancer cell lines.^2,4

According to the previous step in the present study, 7 dilutions of case study solutions were prepared. The human breast cancer cell line MCF-7 was cultured in Dulbecco’s Modified Eagle’s Medium (DMEM), supplemented with 10% fetal bovine serum (FBS) at 37°C with 5% CO₂. Equal numbers of the cells were passaged in 96-well plates (8×10³cells/well) containing 100µL of the medium for 24 hours. One hundred microliters of the different concentrations of the three case study solutions were dispersed in each well and incubated for 24 hours at 37°C with 5% CO₂. Fresh medium (100 µL) containing 0.5 mg/mL of MTT was replaced by the previous media in each well. The growth of the cells was quantified by the ability of the live cells to reduce the MTT Pink dye to a pale violet formazan product. After 4 hours, the formazan product of MTT reduction was dissolved in DMSO, and absorbance was measured using a microplate reader. Effect of AgNP as the percentage of control absorbance of reduced dye was measured at 570 nm as well.