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. 2020 Aug 5;28(12):2677–2690. doi: 10.1016/j.ymthe.2020.08.001

Figure 5.

Figure 5

Combination of In Situ Acute 3WJ-L12 and Subacute iNSC Transplantation Ameliorates Secondary Damage and Promotes Oligodendroglial iNSC Differentiation

(A) Quantification of grafted fGFP-positive iNSCs co-expressing differentiation markers at 56 dpi. Data are mean % of double-positive cells over the total number of GFP-positive cells (±SEM) from n = 3 mice per group. ∗p < 0.05, versus 3WJ-NT/iNSCs. Representative immunofluorescence micrographs depicting engrafted GFP-positive iNSCs co-expressing astroglial marker GFAP (red) or immature oligodendroglial marker Olig2 (red) at 56 dpi (arrow). White rectangular area points to magnification snapshots. (B) Stereological quantification of lesion volumes expressed as % of lesion volume over the volume of selected spinal cord segment. Data represent minimum to maximum, n ≥ 5 mice per group. x axis as per (C) below. (C) Stereological quantification of glial scar volumes measured as GFAP-positive volume at 56 dpi. Data are expressed % of GFAP-positive volume over the volume of selected spinal cord segment. Data represent minimum to maximum volume, n ≥ 4 mice per group. ∗p < 0.05, versus 3WJ-NT/iNSCs. (D) Left: quantification of spared NeuN-positive cells in the ventral horns of experimental groups mice at 56 dpi. NeuN-positive cells were quantified using coronal serial sections of the lesioned area and perilesional area (±1,200 μm). Data are mean numbers per mm2 (±SEM) from n = 5 mice per group. ∗∗p < 0.01, versus 3WJ-NT/iNSCs. Right: representative immunofluorescence micrographs depicting NeuN-positive cells. NeuN in red. (E) Number of iNOS-positive cells and number of CD206-positive cells over the total number of F4/80-positive cells at 56 dpi. Data are mean % of double-positive cells over the total number of F4/80-positive cells (±SEM) from n ≥ 3 mice per group. ∗p < 0.05 and ∗∗p < 0.01, versus 3WJ-NT/iNSCs. Right: representative immunofluorescence micrographs depicting F4/80-positive cells co-expressing iNOS or CD206. x axis of the top graph (iNOS) as per that of the graph below (CD206). Nuclei in (A), (D), and (E) were stained with DAPI (blue). Scale bars, 100 μm. dpi, days post injury.