1. Five‐in‐One: Simultaneous isolation of multiple major liver cell types from livers of normal and NASH mice
- Author
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Andrew J. Murphy, Mark W. Sleeman, Xiping Cheng, Funmilola Adewale, Sun Y. Kim, Ye Zhou, David J. Glass, and Qi Su
- Subjects
Male ,non‐alcoholic steatohepatitis ,Cell type ,Short Communication ,Short Communications ,Fluorescent Antibody Technique ,Cell Separation ,Biology ,liver ,Chronic liver disease ,Immunophenotyping ,cell isolation ,fluorescence‐activated cell sorting ,Mice ,Immune system ,Non-alcoholic Fatty Liver Disease ,medicine ,Animals ,Cluster of differentiation ,Macrophages ,Liver cell ,Endothelial Cells ,RNA ,Cell Biology ,Cell sorting ,medicine.disease ,Cell biology ,Disease Models, Animal ,Hepatocytes ,Hepatic stellate cell ,Molecular Medicine ,Biomarkers - Abstract
NASH is a chronic liver disease that affects 3%–6% of individuals and requires urgent therapeutic developments. Isolating the key cell types in the liver is a necessary step towards understanding their function and roles in disease pathogenesis. However, traditional isolation methods through gradient centrifugation can only collect one or a few cell types simultaneously and pose technical difficulties when applied to NASH livers. Taking advantage of identified cell surface markers from liver single‐cell RNAseq, here we established the combination of gradient centrifugation and antibody‐based cell sorting techniques to isolate five key liver cell types (hepatocytes, endothelial cells, stellate cells, macrophages and other immune cells) from a single mouse liver. This method yielded high purity of each cell type from healthy and NASH livers. Our five‐in‐one protocol simultaneously isolates key liver cell types with high purity under normal and NASH conditions, enabling for systematic and accurate exploratory experiments such as RNA sequencing.
- Published
- 2021