GSM8415439: mouse liver, G0, 10x Genomics; Mus musculus; RNA-Seq
Mice were anesthetized by intraperitoneal injection of 2,2,2-Tribromoethanol, and then perfused with 10mL pre-warmed (37°C) phosphate-buffered saline (PBS) buffer containing 1 mM EGTA through the inferior vena cava. At the start of perfusion, the portal vein was cut to allow blood to be rinsed off. Subsequently, the liver was perfused and digested using 10mL of pre-warmed 1 mg/ml collagenase IV (Sigma‒Aldrich, C5138) in RIPM 1640 medium (Gibco, 21870076). The digested liver tissues were collected into PBS buffer and filtered using a 70μm cell strainer. The resulting cell suspension was centrifuged at 50 × g for 2 minutes at 4 °C, and the cell pellets were resuspended in RIPM 1640 medium supplemented with 1% fetal bovine serum (FBS). To enrich for viable hepatocytes, Percoll (GE, 17-0891-09) diluted in PBS was added to the cell suspension, followed by centrifugation at 50 × g for 2 minutes. The resulting cell pellets were collected, washed once, and resuspended in RIPM 1640 medium supplemented with 1% FBS. 10x Genomics scRNA-seq; cDNA preparation and library construction were conducted using the Single Cell 3' Reagent Kit v3 according to the manufacturer's instructions.