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Updated: Jul 23, 2025

Protocol for Isolation of Primary Human Hepatocytes and Corresponding Major Populations of Non-parenchymal Liver Cells
Published on: March 30, 2016
Protocol to dissociate and isolate wide-diversity single cells by density gradient centrifugation from human
Xianchao Zhou1, Mei Meng1, Yifan Wu2
1State Key Laboratory of Systems Medicine for Cancer, Center for Single-Cell Omics, School of Public Health, Shanghai Jiao Tong University School of Medicine, Shanghai 200025, China.
Abstract:
Single-cell transcriptome sequencing can characterize various cell types in human liver tissue and facilitate understanding of hepatoblastoma heterogeneity. Here, we present a protocol for isolating hepatocytes and immune cells from human hepatoblastoma samples with high viability. We describe steps for tissue processing, enzymatic digestion, Percoll density gradient separation, cell lysis, cell suspension quality control, and scRNA library construction. We then detail sequencing and data analysis. This protocol is applicable to preparing single-cell suspensions from other human liver tissue samples.
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