Fig. 3

DPPSC engraftment, differentiation and revascularisation in skin wounds. a, b Bright field images and corresponding fluorescence images showing tGFP expression (green) in wounds (with silicone rings) at day 5 in mice treated with tGFP⁺ DPPSC (b) but not with PBS (a). Unspecific autofluorescence can be observed in the TRITC channel (upper insets). Scale bars: 5 mm. c, d Immunofluorescence analysis for tGFP (green) in paraffin embedded sections of wounds treated with PBS (c) or DPPSC (d) at day 11. Scale bars: 100 μm. e-g) tGFP (green; e, g) and αSMA (red; f, g) double staining on wound cross-sections reveal double-positive cells (yellow; g) representing DPPSC integrated in a vessel-like structure. Scale bars: 100 μm (e, f), 20 μm (g). h, i CD31 (green) and αSMA (red) immunofluorescence analysis in PBS (h) or DPPSC-treated (i) wounds, showing the presence of CD31⁺ vessels with αSMA coverage (arrowheads). Scale bars: 100 μm. For c-i, nuclei are counterstained with DAPI (blue). j Quantification of αSMA-coated vessels in PBS and DPPSC-treated wounds, showing higher percentage of coverage in DPPSC-treated wounds. *p < 0.05, n = 5 mice/group. k Quantification of the area of αSMA-coated vessels in PBS and DPPSC-treated wounds, showing a larger area of coverage in DPPSC-treated wounds. *p < 0.05, n = 5 mice/group. For j, k, two-tailed Student’s t test was used and results are displayed as mean ± s.e.m