Single cell transcriptome profiling of mouse pancreatic β-lineage cells and hESC-derived β-like cells

While pancreatic β cells have been shown to originate from endocrine progenitors in ductal regions, it remains unclear precisely where β cells emerge and which transcripts define newborn β cells. Here, we used a mouse model “Ins1-GFP;Timer” that provides spatial information during β-cell neogenesis with high temporal resolution. Fluorescent imaging demonstrated that, as expected, some newborn β cells arise close to the ducts; unexpectedly, all the others arise away from the ducts and adjacent to blood vessels. Single-cell RNA-sequencing (scRNA-seq) demonstrated five distinct populations of newborn β cells, confirming the spatial heterogeneity of β-cell neogenesis, and integration analysis with scRNA-seq of hESC-derived β-like cells uncovered transcriptional similarity with the data in mouse β cells. Thus, the combination of time-resolved histological imaging with single-cell transcriptional mapping demonstrated novel features of spatial and transcriptional heterogeneity in β-cell neogenesis, which will lead to a better understanding of β-cell differentiation for future cell therapy. This study characterizes the single-cell transcriptomes of mouse pancreatic β-lineage cells and hESC-derived β-like cells Cellranger aggr was used to combine data from multiple samples (Green1, Green2 and Red); Aggregation file of Green1 and Green2: Green_aggr.barcodes.tsv.gz, Green_aggr.genes.tsv.gz, Green_aggr.matrix.mtx.gz Aggregation file of Green1, Green2 and Red: Green_Red_aggr.barcodes.tsv.gz, Green_Red_aggr.genes.tsv.gz, Green_Red_aggr.matrix.mtx.gz Aggregation file of ES1 and ES2: ES_aggr.barcodes.tsv.gz, ES_aggr.genes.tsv.gz, ES_aggr.matrix.mtx.gz