Fig. 5

Fabrication of scaffold-free SCAP-based spherical calcified constructs. a, b Determination of the optimal requirement including initial seeding cell number and preculture period for SCAP spheroid formation. SCAP were seeded at the indicated initial number per well and were cultured for the indicated period. Representative microscopic images of each scaffold-free SCAP-based spheroids (a). Measurement of the area of the microscopic images of each scaffold-free SCAP-based spheroids (b). c–e Features of scaffold-free SCAP-based spherical calcified constructs. Scaffold-free SCAP-based spheroids were cultured under osteogenic/dentinogenic condition for 4 weeks. Representative inverted (left panel in c) and stereo (right panel in c) microscopic images and 3D (left panel in d) and tomographic (2D, right panel in d) micro-CT images of scaffold-free SCAP-based spheroidal calcified constructs. RT-qPCR analysis shows the expression of RUNX2, BGLAP, and DSPP in scaffold-free SCAP-based calcified constructs. Results show the ratios to the expression of the corresponding 18S ribosomal RNA (18S). nd, no detection (e). a, c, d: Bars = 1 mm (a), 100 μm (c, d). b, e: n = 5 for all groups. Graph bars show the means ± SEM