Assessing Cell Viability and Proliferation

The Vybrant MTT colorimetric assay (Life Technologies, Thermo Fisher) was used to assess cell viability and proliferation in 2D cultures, to determine cell engraftment and localization in 3D cultures and was performed according to manufacturer's instructions. Two-dimensional and 3D-cultured cells were incubated with MTT diluted 1:10 in proliferative medium for 4 h at 37°C. In 2D cultures, the reaction product, formazan, was solubilized in 100 μL of SDS and its concentration was determined by optical density at 550 nm. In 3D cultures, scaffolds were washed with PBS and imaged with a stereomicroscope.^{17 (link)} Formazan-positive hMABs were visible within the scaffolds.