(A) Representative serial sections of spinal cord rostral and caudal to the site of implantation (T9) show that eGFP-mNSCs (green) survive 28 days post-transplantation and migrate towards areas of demyelination compared to PBS treated controls. Nuclei are stained with DAPI (blue) (B) Representative brightfield images of coronal spinal cord sections from eGFP-mNSC (n= 12) or PBS control (n= 13) transplanted animals stained with luxol fast blue (LFB) and counterstained with hematoxylin and eosin (H&E), areas of demyelinated white matter outlined in black. (C) Quantification of demyelination in the ventral white matter of eGFP-mNSC (n= 12) or PBS control (n= 13) transplanted mice revealed significantly (p<0.05) reduced demyelination at sections caudal to the injection site. (D) Quantification of the frequency of adaptive immune cell populations CD4+, CD4+CD25+FoxP3+ and CD8+ T cell subsets within the spinal cord, draining cervical lymph nodes, and spleen of eGFP-mNSC (n= 5) or PBS control (n= 5) injected mice revealed no significant difference in cell populations at day 7 p.t. FACS plots were analyzed and quantified by gating on lymphocytes, excluding forward and side scatter doublets, CD4+ or CD8+ and CD4+CD25+FoxP3+. Sample gating strategy is shown in Supplemental Figure 1. Data represents two independent experiments. 14 days p.t. there was a slight, but significant (*p= 0.0182) decrease in frequency of CD4+ T cells within the spinal cord of mice receiving eGFP-mNSCs 14 days p.t., however no significant difference in the frequency of CD4+ T cells, CD8+ T cells and CD4+CD25+FoxP3+ Tregs in the cLN and spleen 7, 14 or 21 days p.t. or in the SC at day 7 or 21 p.t. For (C), day 28 p.t. demyelinated white matter analysis eGFP-mNSCs (n=12) and control PBS (n=13). Data is presented as average ± SEM and analyzed using one-way ANOVA followed by Tukey’s multiple comparison test. For (D), day 7 p.t. analysis eGFP-mNSCs (n=5) and control PBS (n=5). Data is presented as average ± SEM and analyzed using a two-way ANOVA with a Sidak’s multiple comparisons test.