title: Advanced Method for Isolation of Mouse Hepatocytes, Liver Sinusoidal Endothelial Cells, and Kupffer Cells.
abstract: Separation of pure cell populations from the liver is a prerequisite to study the role of hepatic parenchymal and non-parenchymal cells in liver physiology, pathophysiology, and immunology. Traditional methods for hepatic cell separation usually purify only single cell types from liver specimens. Here, we describe an efficient method that can simultaneously purify populations of hepatocytes (HCs), liver sinusoidal endothelial cells (LSECs), and Kupffer cells (KCs) from a single mouse liver specimen. A liberase-based perfusion technique in combination with a low-speed centrifugation and magnetic-activated cell sorting (MACS) led to the isolation and purification of HCs, KCs, and LSECs with high yields and purity.
authors.affiliation: Department of Infectious Diseases, Union Hospital, Tongji Medical College, Huazhong University of Science and Technology, Wuhan, China.|Institute for Virology, University Hospital of Essen, University of Duisburg-Essen, Hufelandstr. 55, 45122, Essen, Germany.
authors.forename: Jia
authors.initials: J
authors.lastname: Liu
authors.affiliation: Institute for Virology, University Hospital of Essen, University of Duisburg-Essen, Hufelandstr. 55, 45122, Essen, Germany.
authors.forename: Xuan
authors.initials: X
authors.lastname: Huang
authors.affiliation: Department of Gastroenterology and Hepatology, University Hospital of Essen, University of Duisburg-Essen, Essen, Germany.
authors.forename: Melanie
authors.initials: M
authors.lastname: Werner
authors.affiliation: Department of Gastroenterology and Hepatology, University Hospital of Essen, University of Duisburg-Essen, Essen, Germany.
authors.forename: Ruth
authors.initials: R
authors.lastname: Broering
authors.affiliation: Department of Infectious Diseases, Union Hospital, Tongji Medical College, Huazhong University of Science and Technology, Wuhan, China.
authors.forename: Dongliang
authors.initials: D
authors.lastname: Yang
authors.affiliation: Institute for Virology, University Hospital of Essen, University of Duisburg-Essen, Hufelandstr. 55, 45122, Essen, Germany. mengji.lu@uni-due.de.
authors.forename: Mengji
authors.initials: M
authors.lastname: Lu
mesh_terms: D000818:Animals; D015415:Biomarkers; D002469:Cell Separation / Q000379:methods*; D042783:Endothelial Cells / Q000166:cytology*; D005434:Flow Cytometry; D022781:Hepatocytes / Q000166:cytology*; D018189:Immunomagnetic Separation / Q000379:methods; D016130:Immunophenotyping; D007728:Kupffer Cells / Q000166:cytology*; D008099:Liver / Q000166:cytology*; D051379:Mice; D010641:Phenotype
chemical_list: D015415:Biomarkers
keywords: Hepatocytes; Kupffer cells; Liver sinusoidal endothelial cells; Magnetic-activated cell sorting; Technology