Characterization of differentiated CD34⁺ cells at single-cell level. scRNA-seq of ex vivo liquid culture differentiation samples of healthy CD34⁺ cells at day 24 after addition of spCAR or spUTD as control was performed. (A) An overview of the 8259 cells that passed quality control and filtering for subsequent analyses in this study. On the left, UMAP plot showing the 11 clusters that were analyzed and annotated. On the right, UMAP plot showing the distribution of cells from each condition (spUTD or spCAR). (B) Dot plot with the expression of canonical markers. (C) Violin plots of cell markers for mature neutrophils (MPO, PRTN3, ELANE, LTF, and MMP8) and precursors of neutrophils (RUNX1, GATA2, CXCR4, and CEBPA). (D) Violin plots of cell markers for mature monocytes (CD14, CXCL16, IL18, and CD74) and monocyte precursors (CLU, CCL23, VSTM1, and PADI4). (E) UMAP plot showing the expression of KIT and TPSAB1, which are mainly distributed within erythroid precursors. (F) Gene ontology analysis of granulocytes and erythroid precursor clusters corresponding to cells exposed to spUTD and spCAR, respectively, showed pathways that confirm these phenotypes. (G) Expression of the cytokine TGF-β, TNF, and IL-1 receptors families in neutrophils, precursors of neutrophils, monocytes, precursors of monocytes, and erythroid precursors.