Single Cell Isolation from Mouse Liver and Tumor

Prior to flow cytometry analysis, single cell suspensions should be prepared. The method was used as described in the research paper 32 (link). Briefly, after the anesthetization of mice, Hank's buffer without calcium was first injected into the liver through the portal vein. After that, the Hank's buffer with calcium, magnesium and collagenase IV (0.2 mg/mL, Sigma-Aldrich, C5138) was injected into the liver. After separation of the liver and tumor, the tissues were made into small pieces about 1mm³. Mouse tumor dissociation buffer (Miltenyi, 130-096-730) was used to prepare the single cell by using the gentleMACS dissociator (Miltenyi Biotech) followed by filtration through a 70μm cell mesh, lysing erythrocyte, centrifuging and resuspending in Hank's buffer.