Fig. 1
Preliminary evaluation of hNSC survival and efficacy under low- and high-dose transient immunosuppression, and long-term safety study of high-dose immortalized-hNSCs. a) Experimental scheme showing unilateral hNSC transplantation (3 × 105) timeline into the lateral ventricle of n = 5 ALS mice (EXP 1) sacrificed at 20 (n = 2) and 40 (n = 3) dpt under low-dose (15 mg/kg daily) transient (15 days) immunosuppression (left) and representative confocal image of mouse brain showing only cellular debris (right)| huN, green; GFAP, red; DAPI, blue. b) Experimental scheme showing unilateral hNSC transplantation (3 × 105) timeline into the lateral ventricle of n = 12 ALS mice under high-dose (30 mg/kg daily) transient (15 days) immunosuppression (left) (EXP 2) and representative confocal images of mouse brain showing huN+ living cells long-term (right)| huN, green; GFAP, red; DAPI, blue. c) Endurance loss visualization of motor performance decline considering a total of n = 12 mice + 3 × 105 hNSCs (blue line) vs. n = 11 mice + HBSS (red line) (EXP 2). Two-way ANOVA, uncorrected Fisher’s LSD test, Mixed-effect model. Data are expressed as mean values ± SEM. d) Schematic showing the timeline of a bilateral ICV transplantation of 1 × 106 immortalized-hNSCs in n = 3 immunodeficient mice. e) Upper panel: schematic representation of the different brain areas in which huN+ viable cells have been identified.| Lower panel: representative coronal brain sections from Paxinos et al., 2001 and confocal images of surviving huN+ (green) grafts 6 mpt.| huN, green; GFAP, red; DAPI, blue. f) Chart validating both the transplantation and cell biodistribution symmetry after bilateral injections in n = 3 nude mice. Data are expressed as percentage (%) over the total transplanted animals. Drawings on this figure were created with BioRender