Lineage dynamics of pancreatic development at single cell resolution. Mus musculus

A developmental, single-cell transcriptional timecourse analysis was performed on murine pancreas from embryonic days 12, 14, and 17. Whole embryonic murine pancreas tissue was dissociated to single cells, stained with a dead-cell indicator, and subjected to fluorescence activated cell sorting (FACS) to select all live cells. Single cell RNA-sequencing libraries were then subsequently generated for 4,631 cells at E12, 9,028 cells at E14 (comprised of two independent batches), and 4,635 cells at E17. Cells were sequenced at a depth of ~60,000 reads/cell (E14 Batch 1) or ~30,000 reads/cell (E12, E14, and E17 Batch 2). Identification of a novel epithelial progenitor population and multiple novel mesenchymal populations showcases the power of single cell RNA-sequencing to uncover previously uncharacterized cell types, and highlights the dynamic cellular heterogeneity of the developing pancreas. Overall design: Data set 1: Single cell sequencing of embryonic Swiss Webster murine pancreatic tissue using the HiSeq 2500. Live cells were collected from E12.5, E14.5, and E17.5 whole pancreata and prepared using the 10x Single Cell instrument and v1 10X single cell kits according to standard protocol. The first batch consisted of 1 E14.5 litter and was run on 1 well of the 10X instrument. The second batch consisted of 1 well each of E12.5, E14.5, and E17.5 pancreata. Data set 2: Single-cell RNA-sequencing was performed on embryonic Swiss Webster or Fev-Cre;ROSA26mTmG murine pancreatic tissue using the NovaSeq. Live cells were collected from E12.5, E14.5, and E17.5 (Swiss Webster) and E14.5 (Fev-Cre;ROSA26mTmG) whole pancreata and prepared using the 10x Single Cell instrument (10X Genomics) and the v2 10X single cell kits according to manufacturer's instructions.