Measuring Cell Growth via ATP/Luciferase Assay

Growth activity was examined by an ATP/luciferase-based assay (Vialight Plus kit, Lonza, Verviers, Belgium) as described [15 (link)]. Briefly, for measuring growth activity of adherent cells, 10⁴ siRNA-transfected cells were added to each well of a 24-well plate. For determining growth activity in 3D suspension cultures, the cells were incubated in ultra-low attachment 96-well microplates (Corning, Steinfurt, Germany) at a density of 5 × 10³ cells/well. Under both culturing conditions, cells were grown for four days before they were lysed and ATP-measured by the luciferase-based assay in a Sirius luminometer (Berthold). To study the effect of CAF-CM on growth, CAF-CM was added to the medium at a ratio of 1 to 5.