Luminescent cell viability assay (CellTiter-Glo 3D) for cell proliferation and drug response assays

After the cells were grown for the desired amount of time, 100 μL of the CellTiter-Glo 3D reagent was added to each well of the 2D and 3D cultures. The plates were placed on a plate shaker for 5 min, followed by incubation for 30 min at room temperature. For 2D-cultured cells, 100 μL of the solution from each well of the 48-well plate was transferred to a white, clear-bottom 96-well plate after incubation, followed by the addition of the CellTiter-Glo 3D reagent. The luminescence was measured using the PHERAstar Plus microplate reader (BMG Labtech, Offenburg, Germany).

To measure drug sensitivity to chemotherapeutics, the same protocol was followed using the CellTiter-Glo 3D assay. The luminescence readings of Docetaxel- or Rapamycin-treated cells were corrected with the readings of the vehicle control (DMSO equivalents). The cell survival percentage was normalized using the untreated cells as 100% cell survival.