GSM6622231: Proximal; Mus musculus; RNA-Seq
For advanced metastasis, livers were perfused with F4/80-FITC antibody in vivo before dissection. Freshly dissected livers were minced finely and incubated in digestion buffer (1mg/ml collgenase P in Ca2+/Mg2+ containing HBSS) in a shaking incubator at 37 degrees for 30 minutes. Cell extract was further digested with 0.25% trypsin-EDTA at 37 degrees for 5 minutes. After removal of RBCs, cells were then resuspended in PBS + 0.5% BSA and stained with viability dye and CD45, CD11b, F4/80-APC antibodies. Liver F4/80+ cells were then isolated using FACS. Library construction was performed according to the manufacter's instructions (single cell 3.1' v3 protocol, 10x Genomics).