Figure 5

Evaluation of myogenic differentiation and adipogenic differentiation in STRO-1 ⁺ /c-Kit ⁺ /CD34 ⁻ hDPSCs and STRO-1 ⁺ /c-Kit ⁺ /CD34 ⁺ hDPSCs. A: Confocal analysis of myogenic differentiation of STRO-1+/c-Kit+/CD34⁻ hDPSCs and STRO-1⁺/c-Kit⁺/CD34⁺ hDPSCs directly co-cultured with C2C12 mouse myoblasts. Double immunofluorescence labeling shows signals from anti-hMit (green) and anti-myosin (red) and from anti-hNu (green) and anti-myosin (red), respectively. DAPI staining is shown in blue. Bar: 10 μm. B: Evaluation of adipogenic commitment in STRO-1⁺/c-Kit⁺/CD34⁻ hDPSCs and STRO-1⁺/c-Kit⁺/CD34⁺ hDPSCs. On the top, histological staining with Oil Red O showing adipogenic differentiation of STRO-1⁺/c-Kit⁺/CD34⁻ hDPSCs and STRO-1⁺/c-Kit⁺/CD34⁺ hDPSCs. Cells were counterstained with haematoxylin. Bar: 10 μm. Below, AdipoRed assay performed on both hDPSCs subpopulations, evaluated for lipid droplets formation by confocal microscopy analysis. Differentiated STRO-1⁺/c-Kit⁺/CD34⁻ hDPSCs and STRO-1⁺/c-Kit⁺/CD34⁺ hDPSCs show a green fluorescent staining. Bar: 10 μm. At the bottom, quantification of triglycerides (RFU, relative fluorescent unit) carried out by fluorimeter analysis. ***p<0,001 diff STRO-1⁺/c-Kit⁺/CD34⁻ hDPSCs vs undiff STRO-1⁺/c-Kit⁺/CD34⁻ hDPSCs; ^§§§ p<0,001 diff STRO-1⁺/c-Kit⁺/CD34⁺ hDPSCs vs undiff STRO-1⁺/c-Kit⁺/CD34⁺ hDPSCs, ANOVA test followed by Newman-Keuls post-hoc test.