Fig. 5: CFU-M progenitors have endothelial-macrophage plasticity and vasculogenic capacity in vivo.

a Schematic of 1° transfer of GFP⁺ aortic progenitors in hindlimb muscle after hindlimb ischemia surgery with 14 d follow-up. b Laser Doppler perfusion images of mice on day 0 and 14 after ischemia surgery and receiving cell-free control (above) or progenitors (Prog; below). Graph shows results of follow-up over 14 d (n = 6 mice). Data was analyzed using mixed effects two-way ANOVA (p < 0.0001 for time, p = 0.03 for group and p = 0.005 for time x group) with Sidak’s multiple comparisons test (#p = 0.0009 for progenitor vs control). c Fluorescence-minus-one (FMO) control staining used to analyze the composition of cells produced by engrafted GFP⁺ progenitors after transfer into ischemic hindlimb muscle. GFP, green fluorescent protein. d Flow cytometry plots and graph show the cells produced by donor cells in recipient muscle (n = 6 mice). Green histogram, sample; dotted histogram, FMO control. Data was analyzed using repeated measures one-way ANOVA with Tukey’s multiple comparisons test (*p = 0.02 vs Mϕ). Mϕ, macrophages; EC, endothelial cells. e Confocal microscopy of immunolabeled recipient ischemic muscle shows neovessels lined by GFP⁺CD31⁺ endothelial cells (arrows) with cluster of GFP⁺CD68⁺ macrophages (arrowhead) (above) and perfused with host TER-119⁺ erythrocytes (below). L, lumen. f Schematic of 2° transfer of GFP⁺ progenitors into hindlimb muscle after hindlimb ischemia surgery. M/C, Methylcellulose. g Flow cytometry plots and graph show cells produced by donor cells after 2° transfer (n = 3 mice; repeated measures one-way ANOVA with Tukey’s multiple comparisons test; *p = 0.02 for Mϕ and p = 0.04 for EC vs Prog). h Schematic of 1° transfer of GFP⁺ aortic progenitors in hindlimb muscle after induction of hindlimb ischemia with 56 d follow-up. i Flow cytometry plots and graph show cells produced by donor cells 8 w after 1° transfer (n = 3 mice; repeated measures one-way ANOVA with Tukey’s multiple comparisons test; *p = 0.02 for Mϕ and †p = 0.008 for Prog vs EC). Data summarized as mean ± SD. Scale bar, 20 µm. Also see Supplementary Fig. 10. Source data are provided as a Source Data file. Figure 5 panels (a), (f) and (h) created with BioRender.com released under a Creative Commons Attribution-Non-commercial-No Derivs 4.0 International license.