Single-cell RNA sequencing analysis of PDOs from 4 non-EoE patients. PDOs from 4 non-EoE patients were treated with IL13 (10 ng/mL) or vehicle (phosphate-buffered saline) from day 7 to day 11, and harvested and dissociated for single-cell RNA sequencing analysis on day 11. (A) Seurat’s Uniform Manifold Approximation and Projection (UMAP) visualization of 4 classified cell groups, basal-1, basal-2, suprabasal, and superficial, within PDOs from non-EoE controls grown with or without IL13 (10 ng/mL). (B) Average normalized gene expression values of MKI67, CD73, basal markers (TP63, NGFR), and differentiated markers (IVL, KRT13, FLG) in each cell cluster. Circle size reflects the percentage of cells with a nonzero expression level for the indicated genes. Color intensity reflects the average expression level across all cells within each cluster. (C) Expression z-scores for the top 8 up-regulated genes in each cluster. (D) The proportions of each cell cluster were compared in PDOs (n = 4) with IL13 or vehicle. (E–G) The normalized expression of (E) IVL and FLG, (F) SOX2, and (G) CD73 in nontreated (NT) PDOs was compared with that in PDOs treated with IL13 for each cluster. ∗∗∗∗P < .0001, Wilcoxon signed-rank test. (H) GO analysis of differentially expressed gene profiles of CD73+CD104+ cells, compared with CD73-CD104+ cells in NT PDOs.