Single-cell characterization of macrophages in fibrotic and regenerative microenvironments. Single-cell characterization of macrophages in fibrotic and.

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Single-cell characterization of macrophages in fibrotic and regenerative microenvironments. Single-cell characterization of macrophages in fibrotic and regenerative microenvironments. (A) Experimental overview. A virtual aggregate of macrophages in fibrosis and regeneration generated from scRNAseq after sorting of F4/80hi+CD64+. Cells were isolated from murine volumetric muscle injuries at 1 week, treatment with biomaterials UBM (regenerative, IL-4 tissue environment), synthetic (fibrotic, IL-17–rich environment), or saline (wound control). (B) Heatmap of differentially expressed genes. Up to 200 cells per cluster are shown, ordered by cluster, with the top 10 differentially expressed genes. Functionally relevant genes from terminal clusters are annotated. (C) Dimensional reduction projection of cells onto two dimensions using UMAP. Cells are colored by experimental biomaterial condition (top) and computationally determined cluster (bottom). (D) Summary of cluster differentiation trajectories, composition by experimental origin, markers, and biological functions generated by bioinformatics analysis. (E) Flow cytometry strategy informed by computationally determined markers including CD9, CD301b, and MHCII differentiating in vivo macrophage subsets from UBM or synthetic biomaterial. Subsets are colored equivalent to computational clusters, back-gated into tSNE projection. Sven D. Sommerfeld et al. Sci. Immunol. 2019;4:eaax4783 Copyright © 2019 The Authors, some rights reserved; exclusive licensee American Association for the Advancement of Science. No claim to original U.S. Government Works