Extended Data Fig. 4: Muscle regeneration with angiogenesis and innervation.
From: Bioinstructive scaffolds enhance stem cell engraftment for functional tissue regeneration
a, Representative images (left) and quantification (right) of self-renewed transplanted cells (RFP+Pax7+) within the region of regenerated myofibers (n = 6 biologically independent experiments). Scale bar, 10 μm. b, Representative FACS plots of endothelial cell isolation. c, A representative image demonstrating the purity of isolated endothelial cells. Scale bar, 20 μm. d–f, Representative images (d) and quantification of the number (e) and length (f) of tube formation by endothelial cells treated with PBS, NCFGF(E), or NCFGF(E)/NCIGF(L) constructs (n = 4 independent synthesis experiments with endothelial cells pooled from three mice). Scale bar, 100 μm. g, h, Representative images (g) and quantification (h) of endothelial cell infiltration into different scaffolds transplanted into VML defects without MuSCs 2-week post-transplantation (n = 4 biologically independent experiments). Dashed yellow line indicates the edge between host tissue (left of the line) and scaffold (right of the line). Scale bar, 100 μm. i–k, Quantification of the density of endothelial cells (i), functional capillaries (j), and the number of capillaries per myofiber within the region of donor-derived fibers 6-week post-transplantation of RFP+ MuSCs seeded onto different bioconstructs (n = 6 biologically independent experiments). l, Representative images (left) and quantifications (right) of neovessels surrounded by pericytes (PDGFRβ+) and vascular smooth muscle cells (α-SMA+) within the region of RFP+ regenerated myofibers (n = 3 biologically independent experiments). Scale bar, 10 μm. m, Representative images (left) and quantification (right) of Neuro2a neuroblast differentiation after treatment with PBS, NCFGF(E), or NCFGF(E)/NCIGF(L) constructs. The arrows indicate neurites of differentiated Neuro2a cells (n = 3 independent synthesis experiments). Scale bar, 25 μm. Data in a, e, f, h-m are presented as the mean ± SEM. P values in a, e, f, h-k, m were determined by one-way ANOVA. *P ≤ 0.05, **P ≤ 0.01, ***P ≤ 0.001, ****P ≤ 0.0001; ns, not significant.
