Modifications Required for ATP and Caspase Detection Assays Applied to 3D Cell Spheroids

This scientific poster investigates whether cell viability and apoptosis assays designed for 2D monolayers of cells would work effectively with 3D spheroid models. Cell viability assay reagents that measure ATP contain detergent to lyse cells and release ATP. Some commercial reagents have been found to have a low recovery of ATP from spheroids compared to acid extraction accepted as the gold standard. Reformulation of ATP assay reagent to contain higher amounts of detergent, increasing mixing time using a plate shaker, and using a longer incubation time in the presence of lytic reagents resulted in improvements in extraction of ATP from large 3D cell spheroids.