High-resolution single cell RNAseq of ALK3-expressing human pancreatic ductal cells

Purpose: We demonstrate here that human pancreatic ductal cells within the exocrine pancreas have multiple cell type populations. Flow cytometry and live-cell sorting of ALK3bright+ cells enabled transcriptomics analysis at the single cell level. Methods: Live CD90-/ALK3bright+ cells from 3 non-diabetic human donors were sorted and cryopreserved. After cryopreservation and dead cell removal, we performed single-cell cDNA library construction using the 10X Genomics microfluidics platform, utilizing the chromium single cell 3' v2 chemistry kit. Single cell libraries were sequenced on an Illumina sequencing platform with 10X Genomics recommended parameters. After sequencing, data was converted to filtered read counts via the 10X Genomics Cellranger v3.0.2 software. Filtered gene counts were then utilized for downstream analysis using the Seurat scRNAseq analysis package. Conclusions: Our research presents the first detailed analysis at the single-cell level of ALK3-expressing ductal cells isolated from the human exocrine pancreas. This study reveals heterogeneity across the human ductal compartment, with certain sub-populations of ductal cells exhibiting progenitor-like characteristics. These data also demonstrate plasticity across the exocrine pancreas, which is donor-dependent. Overall, this study furthers our understanding of pancreatic biology at the single cell level. Overall design: Single cell RNA sequencing analysis of human exocrine pancreas-derived CD90-/ALK3bright+ cells.