Fig. 5: Unsupervised characterization of identified clusters from S6w1 scRNAseq dataset.

a UMAP projection of combined two samples of S6w1 islets from a healthy iPSC line representing annotated islet cell population guided through islet development feature expression without reference dataset supervision. Samples were analyzed using 10X genomics software Loupe Browser v7.0.01. Clusters identified and annotated for frequency distribution as- SC- β (27%), SC-α (10%), SC-δ (1%), SC-γ (<1%), ductal (9%), SC-EC (7%), SC-polyhormonal (8%), Endothelial cells (16%), Endocrine cells (3%), non-endocrine cells (13%). b Comparative cluster identification of each sample obtained from 0.1 L and scale-up 0.5 L generated S6w1 islets to show in UMAP projection and liberality of differentiation without variability. c Feature plots of SC- β cell cluster highlighting functional β cell marker expression. Scale represents log2FC expression (0–6). Color highlight as pale yellow: downregulation and crimson red: upregulation. d Feature plots of selected endocrine clusters representing expression for islet cell differentiation and exocrine markers. Scale represents log2FC expression (0–6). Color highlight as pale yellow: downregulation and crimson red: upregulation. e Heat map plot with Manhattan neighbor distribution clustering representing differentially expressed genes associated with islet developmental pathway between identified and unsupervised annotated clusters. Scale represents log2FC (−1.5 to 1.5), where red color shows upregulation and blue corresponds to downregulation of selected gene expression. f, Violin plots comparing distribution of β and α cell makers for functional maturation gene expression between SC- β, -α, -PH, and endocrine cells. Each plot shows median gene expression distribution with interquartile range for 27–75%. g Dot plot representing differentially expressed genes for glycolysis and TCA metabolism, h Dot plot representing differentially expressed genes for insulin secretion, i Dot plot representing differentially expressed genes for exocytosis, and j Dot plot representing differentially expressed genes for disallowed genes between SC- β, -α, -δ -γ, - ε, -PH, -EC, and ductal cells. k Feature plots of endocrine clusters representing disallowed and off-target pluripotency associated genes expression. Scale represents log2FC expression (0–13). Color highlight as pale yellow: least expression and crimson red: upregulation, gray represents absence of gene expression.