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

While pancreatic ß cells have been shown to originate from endocrine progenitors in ductal regions, it remains unclear precisely where ß cells emerge and which transcripts define newborn ß cells. Here, we used a mouse model “Ins1-GFP;Timer” that provides spatial information during ß-cell neogenesis with high temporal resolution. Fluorescent imaging demonstrated that, as expected, some newborn ß 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 ß cells, confirming the spatial heterogeneity of ß-cell neogenesis, and integration analysis with scRNA-seq of hESC-derived ß-like cells uncovered transcriptional similarity with the data in mouse ß cells. Thus, the combination of time-resolved histological imaging with single-cell transcriptional mapping demonstrated novel features of spatial and transcriptional heterogeneity in ß-cell neogenesis, which will lead to a better understanding of ß-cell differentiation for future cell therapy. Overall design: This study characterizes the single-cell transcriptomes of mouse pancreatic ß-lineage cells and hESC-derived ß-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