Cell adhesion Assay

Cells were seeded at a concentration of 5X10⁴ in 96 well vitronectin coated plates (5 μg/ml). The cells were allowed to adhere for 1 hour at 37°C. Non-adherent cells were removed and the plates were washed twice with warm PBS. The number of attached cells was determined by MTT (3-(4, 5-dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide) assay. MTT (5mg/ml) was suspended in PBS and 10% MTT solution was prepared in serum free medium. The MTT solution was added to the adherent cells and incubated at 37°C for 2 hours. The MTT solution was gently removed and 200 μl DMSO was then added to each well, and incubated for 15 minutes at 37°C. The absorbance was measured using a VersaMax microtiter plate reader at 600 nm.