Fig. 6: Dysfunctional mitochondria released from iPSC-derived microglia (YZ1 line from a healthy donor) induced neurodegeneration and neuroinflammation progression 24 hours after contusion injury.

a Schematic representation of the experimental design. b Representative images of Tuj1 network density in tricultures with iPSC-derived microglia. c Quantification of the Tuj1 neuronal network density in control and P110 treated groups of NAMc. d Mitochondria fission associated pDRP1 and FIS1 proteins expression in control and P110 treated NAM cultures. Proteins were isolated from entire scaffolds without separating the injured area from the penumbra. e Inflammatory cytokines secretion post-injury in YZ1 triculture groups. f Gene set enrichment analysis revealed distinct expression modules associated with M1 - neuronal systems (negatively associated with a contusion in CTRL; rescued with P110); M5 - replication and repair (positively associated with a contusion in CTRL and P110); M6 - interferon signaling (positively associated with a contusion in CTRL; unchanged in P110). g Critical regulators of molecular mechanisms associated with M1, M5, and M6 modules. The full map of 7 detected modules and corresponding pathways can be found in Supplementary Fig. 25, 26, and 29. Data presented in (c, d, e) mean ± SEM of three independent experiments with n = 3 for sham groups and n = 4 scaffolds for injury groups. Data presented in (f, g) were analyzed from mRNA sequencing data collected from n = 6 for sham groups and n = 6 scaffolds for injury groups from two independent experiments. *, **, *** indicates a significant difference (p < 0.05, 0.01, 0.001 respectively; two-way ANOVA (analysis of variance) or one-way ANOVA between control and experimental groups) (with Tukey’s post-hoc test). Experiments were replicated at least three times. Scale bar: 50 μm in b, f, h; 10 μm in j.