Figure 2
3D ACC constructs express key biomarkers in 3D while proliferating over time in vitro. Immunofluorescence staining of key ACC-relevant biomarkers, SF-1, Melan-A, inhibin α, IGF2, and β-catenin both (a–e) 3D construct cultures and (f–j) 2D cultures; insets depict stains without DAPI overlay. (m) Quantification of ATP activity in 3D ACC constructs over time. Statistical significance: *p < 0.05. (k) 3D constructs were maintained for up to 17 days. (l) Brightfield imaging demonstrates 3D construct cell density. (m) LIVE/DEAD staining shows largely viable cells within 3D constructs over the 17 days of culture. Green—calcein AM-stained viable cells; Red—ethidium homodimer-1-stained dead cell nuclei. Scale bars in panels (a–m): 100 µm; (n–o): 200 µm.
