Fig. 1. In vitro and in vivo characterization of GDNF-hNSPCs. (A) Compared with Mock-hNSPCs (DAPI+, blue), many of GDNF-hNSPCs express GDNF (green). (B) An ELISA of GDNF in Mock- and GDNF-CM. (C, D) Representative images of Mock- and GDNF-hNSPCs stained for differentiation markers and quantification of the percentages of cells that expressed these markers. DAPI+ Cells (blue) express nestin (red), GFAP (green), Tuj1 (red), PDGFR (green), and O4 (red). (E, F) Representative images of CM-treated SH-SY5Y cells and quantification of average neurite length under different experimental conditions. (G) hNuc+ GDNF–hNSPCs (red) implanted into the injured spinal cord show robust engraftment and an extensive distribution throughout the lesion and adjacent areas. The boxed area is shown at high magnification in the right panel. The asterisk indicates the lesion epicenter and arrows indicate the cell transplantation sites rostral and caudal to the lesion epicenter. (H, I) Representative images of Mock- and GDNF-hNSPCs stained for differentiation markers in vivo and quantification of the percentages of cells that expressed these markers. Confocal images show that hNSPCs positive for the human cell markers Ku80, STEM101, and STEM121 colocalize with differentiation markers (green). (J) Engrafted STEM121+ GDNF-hNSPCs (red) express GDNF (green). Scale bars: 50 μm in A, D; 100 μm in E; 1 mm in G; and 30 μm in I, J. Data represent the means±SEM. *p<0.05 vs. Mock-hNSPCs in C, H; *p<0.05 vs. vehicle, ***p<0.001 vs. vehicle, ##p<0.01 vs. Mock-hNSPCs in F.
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