Single-cell analysis of whole hepatic non-parenchymal cells from normal and CCl4-injured mice

This study used 10X Genomics, single-cell RNA-sequencing to examine the heterogeneity of liver non-parenchymal cell types present in the normal and CCl4-mediated fibrotic mice. Two male mice of 8-week old were injected i.p with olive oil and CCl4 independently twice a week for a total 6 weeks. Then, liver non-parenchymal cells were obtained by a two-step collagenase perfusion method. Hepatocytes were elimanated by three times of low-speed centrifugation at 50g for three minutes and were not included in further analysis of this study. Single-cell suspensions were then loaded on a Chromium Single Cell Instrument (10X Genomics) to generate single-cell GEMs. After reverse transcription, libraries were prepared using version 3 Chromium Single-cell 3' Library, Gel Bead & Mutiplex Kit (10X Genomics). Sequencing was performed on illumina HiSeq 3000.