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Fig. 5 | Stem Cell Research & Therapy

Fig. 5

From: Three-dimensional cultured mesenchymal stem cells enhance repair of ischemic stroke through inhibition of microglia

Fig. 5

Transcriptomic changes in microglia with MSC administration. A A schematic diagram depicting sample preparation for RNA-seq. CD45lowCD11b+ microglia were isolated from the brain of Sham rats or the ischemic hemisphere 2 days after cell infusion. Total RNA was extracted from isolated microglia for RNA-seq analysis. B PCA (a) of microglia population sampled from sham rats, PBS-, 2D MSC-, or 3D MSC-treated rats using PC1 and PC2 suggests clear separation of sham microglia and 3D MSC-treated microglia from PBS and 2D MSC-treated microglia. Venn diagram (b) showing the interactions among DEGs between Sham with PBS, 2D, or 3D MSC-treated samples. C Heatmap (a) showing the expression profiles of 341 genes which participate in the immune system. Gene Ontology (GO) biological process analysis (b) for the 530 downregulated genes by 3D MSC infusion compared with PBS and 2D MSC infusion showed genes involved in biological processes including cellular response to lipopolysaccharide, inflammatory responses, immune response, and immune system process. Gene set enrichment analysis (GSEA) (c) showing positive enrichment in transcriptome with treatment of PBS and 2D MSCs for immune response gene set. Heatmap (d) showing the expression profiles of genes belonging to proinflammatory mediators. D The expression of Mincle based on RNA-seq data. FPKM, fragments per kilobase of transcript per million mapped reads. E The expression of Mincle was validated with qRT-PCR. *p < 0.05 compared with PBS group. F The expression of Mincle was determined in LPS-activated microglia with treatment of 2D or 3D MSCs conditioned media. *p < 0.05 compared with LPS group

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