Single cell transcriptomics of partial hepatocyte reprogramming

This study analyzes hepatocytes with Notch-induced reprogramming by single cell RNAseq compared to normal hepatocytes and biliary epithelial cells (BECs) in the normal liver. Three-weeks post-hydrodynamic tail vein injection (HDTVi) of our TTR>NICD-CAG>tdT Piggy-Bac transposon in an 8-week-old C57Bl/6 mouse, liver parenchymal cells were isolated and sorted by Fluorescence Activated Cell Sorting (FACS). tdT- hepatocytes, tdT+ reprogramming cells, and EpCAM+ BECs were sorted and subsequently mixed in a 1:2:1 ratio, respectively. This process was repeated in a second mouse and both samples were analyzed for their transcriptional profile by scRNAseq.