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Figure 9: Mesenchymal differentiation and neurosphere-like structure formation of HF (c, e, g, i) and SD NCSC cultures (d, f, h, j) at P3. (a) Osteogenic differentiation, 14 days, qPCR analysis. (b) Adipogenic differentiation, 14 days, qPCR analysis. The expression of all target genes was normalised to TBP gene expression levels and then to the undifferentiated control cells. The HF NCSC and SD NCSC cultures from five donors were analysed. (c, d) Osteogenic differentiation, 10 days, stain with BCIP®/NBT Liquid Substrate System for ALP activity (dark blue). (e, f) Osteogenic differentiation, 21 days, Alizarin Red S stain for mineralized extracellular matrix deposits (red). (g, h) Adipogenic differentiation, Oil Red O stain for neutral lipid vacuoles (red) and Romanowsky-Giemsa counterstain. (i, j) Neurosphere-like structures, phase-contrast microscopy. (c–h) Scale bar: 100 μm. (i, j) Scale bar: 50 μm.