Fig. 2. Grafted GDNF-hNSPCs enhance neurite outgrowth, axonal extension, and myelination in the injured spinal cord. (A) Multiple NF+ neuronal processes (green) extend over engrafted hNuc+ human cells (red). Compared with Mock-hNSPCs, GDNF-hNSPCs significantly enhance neurite extension and host axonal sprouting. Arrows indicate hNuc+ donor-derived cells (red) and arrowheads indicate NF+ neuronal fibers (green). (B) Under high magnification, many NF+ neuronal processes (red) are co-labeled with an anti-MBP antibody (green). Arrowheads indicate co-labeled myelinated neuronal fibers (yellow or orange). (C) The intensity of MBP staining in engrafted and adjacent areas is markedly higher in the GDNF-hNSPC-treated group than in the Mock-hNSPC-treated group. Arrows indicate hNuc+ donor-derived cells (red) and arrowheads indicate MBP+ myelinated neuronal fibers (green). (D) Representative images of LFB staining (blue) in the lesion and adjacent areas of the spinal cord. Dashed lines indicate demyelinated areas. (E) Quantification of the demyelinated volume in the different experimental groups. (F, G) Representative electron microscopic images of axially sectioned spinal cords (T8~T10) and quantification of G-ratio. (H, I) Representative images of Schwann/2E immunostaining (green) in the injured spinal cords and quantification of immunostaining density in the different experimental groups. Scale bars: 1 mm in A, C, D; 100 μm in B; 1 μm in F, and 500 μm in H. Data represent the means±SEM. *p<0.05 vs. vehicle, #p<0.01 vs. Mock-hNSPCs.
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