Liver cells were sorted from mixed, hepatocyte, and non-parenchymal cell
fractions on an Aria Fusion I using a 100 μm nozzle. Cells from the HCC
samples were not fractionated and were sorted directly after tissue digestion.
Zombie Green (Biolegend) was used as a viability dye. Cells were stained with
human specific antibodies against CD45 (Biolegend), PECAM1 (Biolegend), CD34
(Biolegend), CLEC4G (R&D systems), ASGR1 (BD Biosciences), EPCAM
(R&D systems), and TROP2 (Biolegend). Organoids were stained with
antibodies against EPCAM and TROP2. For the humanized mouse samples, cells were
stained either with antibodies against ASGR1 and PECAM1 or human HLA-ABC (BD
Biosciences) and mouse H2-Kb (BD Biosciences). Viable cells were sorted in an
unbiased fashion or from specific populations based on the expression of markers
into the wells of 384 well plates containing lysis buffer.
fractions on an Aria Fusion I using a 100 μm nozzle. Cells from the HCC
samples were not fractionated and were sorted directly after tissue digestion.
Zombie Green (Biolegend) was used as a viability dye. Cells were stained with
human specific antibodies against CD45 (Biolegend), PECAM1 (Biolegend), CD34
(Biolegend), CLEC4G (R&D systems), ASGR1 (BD Biosciences), EPCAM
(R&D systems), and TROP2 (Biolegend). Organoids were stained with
antibodies against EPCAM and TROP2. For the humanized mouse samples, cells were
stained either with antibodies against ASGR1 and PECAM1 or human HLA-ABC (BD
Biosciences) and mouse H2-Kb (BD Biosciences). Viable cells were sorted in an
unbiased fashion or from specific populations based on the expression of markers
into the wells of 384 well plates containing lysis buffer.