Celltiter glo ctg 3d cell viability assay

For comparison of anti-growth activity, a CellTiter-Glo (CTG) 3D cell viability assay (Promega, G9682) was used. Cells (1 × 10⁴ cells per well) were seeded (using the same media) in ultra-low attachment surface 96-well format plates (Corning Costar #3474). On the day after plating, cells were treated with a 7 point three-fold dilution series of indicated compounds (200 μL final volume per well), and cell viability was monitored at 1, 3, 5 days according to the manufacturer’s recommended instructions, after which 50 μL of CellTiter-Glo reagent was added. Samples were then vigorously mixed, covered, and placed on a plate shaker for 20 min to ensure complete cell lysis prior to assessment of luminescent signal.

HCT 116 cells were seeded in 96-well round bottom ultra-low attachment plates (1000 cells per well) and allowed to form spheroids for 72 h. All drugs were dissolved in DMSO, nanoparticles were suspended in DDW, and added to the wells. Untreated cells (control) were used to establish 100% viability. The effect on cell viability was measured with Promega^® CellTiter-Glo^® (CTG) 3D cell viability assay.