Isolation of Hepatic Cells and Immune Cells

Hepatocellular carcinoma mouse liver and tumour tissues were harvested and disaggregated by mechanic mincing and enzymatic digestion with 0.1 mg·mL^–1 DNAse (Roche), 1 mg·mL^–1 type IV collagenase (Invitrogen), and 0.5% hyaluronic acid (Solarbio) for 1 h at 37 °C, then squeezed through a 200‐mesh stainless steel strainer, and the cell suspension was collected. After a brief centrifugation at 100g for 1 min, the liver or tumour cells were at the bottom of tube, and mononuclear cells were isolated from the upper suspensions using Percoll gradient centrifugation as described before [25 (link)]. After washing with PBS, all types of cells were used for flow cytometry analysis.