2.2.3. CyQUANT™ DNA quantification for cell proliferation

The CyQUANT™ cell proliferation assay kit (Thermo fisher scientific) was used to quantify DNA content of 3D cell encapsulated hydrogels. Frozen hydrogel samples (after aspiration of medium and washing with PBS), harvested at different time points, were stored in a −80 °C freezer. The samples were thawed and lysed by addition of a buffer containing the CyQUANT™ GR dye, and incubated at room temperature for 1 h before measuring the fluorescence intensity of the supernatant (1 mL CyQUANT™ GR/cell lysis working solution contains 2.5 μL of the CyQUANT™ GR dye stock solution (400 X in DMSO) in nuclease-free distilled water). 200 μL of gel lysis solution was transferred into a black 96-well plate (Corning). The samples were measured using a fluorescence reader at an excitation of 480 nm and emission of 520 nm.