Evaluating Cell Proliferation and Viability

The (3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide) (MTT) assay was used to evaluate proliferation of viable cells, as described previously^{32 (link)}. The absorbance was read using a microplate (ELISA) reader mQuant spectrophotometer (BIO-TEK Instruments, Inc., Winooski, VT, USA) at 570 nm. Cell viability was assessed using the trypan blue exclusion assay as described elsewhere^{32 (link)}. Cell counting at 48 hrs was performed in triplicate. For colony-formation assay, cells were plated in 24-well plates in 300 µl of MethoCult methylcellulose solution (Sigma, St Louis, MO) at a concentration of 500 cells/well and were incubated for 2 weeks until colony formation was visible. Colonies were counted after staining with p-iodonitrotetrazolium violet (Sigma, St Louis, MO).