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. 2021 Nov 18;6:75. doi: 10.1038/s41536-021-00185-5

Fig. 5. cECFC treatment attenuates glial activation in FGR brains.

Fig. 5

Representative labelling of microglial cells (Iba-1; black) in the parietal cortex. A NG brains display typical ramified microglial morphology, with even distribution, small cell bodies, and extended processes (see insert). FGR demonstrate significant activation of microglia with dense cellular bodies, retraction and thickening of processes, and a high degree of cellular overlap. cECFC-treated animals displayed morphology comparable to NG. B Microglia were assigned as resting or ramified based on well-established morphology associated with activation state of glial cells. C FGR demonstrated significantly higher total number of Iba-1 positive cells and activated microglia (D). Similar trends were observed in the white matter of FGR brain (see Supp. Fig. 3). E Astrocytes (GFAP; white) showed evident alterations in morphology. NG displayed typical star like cells with long process extensions. FGR displayed activated astrocyte morphology, with retracted processes and enlargement of cell bodies. cECFC-treated animals displayed similar morphology to NG. F cECFC treatment reduced the number of astrocytes in the cortex toward levels comparable to other groups. G Treatment with cECFCs reduced astrocyte activation as assessed with GFAP-positive stained area. H Heat map of porcine inflammatory cytokine and receptors arrays demonstrates altered expression of pro- and anti-inflammatory mediators in the cortex of FGR relative to NG. FGR + cECFC showed alterations in inflammatory genes relative to NG and untreated FGR (n = 8 for all groups, pooled samples of n = 4 for each group per array). Expression of well-characterised pro-inflammatory I and anti-inflammatory K genes relative to NG. FGR + cECFC displayed down-regulation of pro-inflammatory (J) and up-regulation of key anti-inflammatory genes (L) when compared with untreated FGR. All values are expressed as mean ± SEM (minimum n = 6 for all groups). Two-way ANOVA with Tukey post-hoc test (*p < 0.05, **p < 0.01, ***p < 0.001, ****p < 0.0001) (Scale bars: 50 µm).