title: All-In-One: Advanced preparation of Human Parenchymal and Non-Parenchymal Liver Cells.
abstract: OBJECTIVE
  Liver cells are key players in innate immunity. Thus, studying primary isolated liver cells is necessary for determining their role in liver physiology and pathophysiology. In particular, the quantity and quality of isolated cells are crucial to their function. Our aim was to isolate a large quantity of high-quality human parenchymal and non-parenchymal cells from a single liver specimen.
  
  
  METHODS
  Hepatocytes, Kupffer cells, liver sinusoidal endothelial cells, and stellate cells were isolated from liver tissues by collagenase perfusion in combination with low-speed centrifugation, density gradient centrifugation, and magnetic-activated cell sorting. The purity and functionality of cultured cell populations were controlled by determining their morphology, discriminative cell marker expression, and functional activity.
  
  
  RESULTS
  Cell preparation yielded the following cell counts per gram of liver tissue: 2.0 ± 0.4 × 10(7) hepatocytes, 1.8 ± 0.5 × 10(6 )Kupffer cells, 4.3 ± 1.9 × 10(5) liver sinusoidal endothelial cells, and 3.2 ± 0.5 × 10(5) stellate cells. Hepatocytes were identified by albumin (95.5 ± 1.7%) and exhibited time-dependent activity of cytochrome P450 enzymes. Kupffer cells expressed CD68 (94.5 ± 1.2%) and exhibited phagocytic activity, as determined with 1 μm latex beads. Endothelial cells were CD146(+) (97.8 ± 1.1%) and exhibited efficient uptake of acetylated low-density lipoprotein. Hepatic stellate cells were identified by the expression of α-smooth muscle actin (97.1 ± 1.5%). These cells further exhibited retinol (vitamin A)-mediated autofluorescence.
  
  
  CONCLUSIONS
  Our isolation procedure for primary parenchymal and non-parenchymal liver cells resulted in cell populations of high purity and quality, with retained physiological functionality in vitro. Thus, this system may provide a valuable tool for determining liver function and disease.
authors.affiliation: Department of Gastroenterology and Hepatology, University Hospital of Essen, University Duisburg-Essen, Hufelandstr. 55, 45147, Essen, Germany.
authors.forename: Melanie
authors.initials: M
authors.lastname: Werner
authors.affiliation: Department of Gastroenterology and Hepatology, University Hospital of Essen, University Duisburg-Essen, Hufelandstr. 55, 45147, Essen, Germany.
authors.forename: Sabrina
authors.initials: S
authors.lastname: Driftmann
authors.affiliation: Department of Gastroenterology and Hepatology, University Hospital of Essen, University Duisburg-Essen, Hufelandstr. 55, 45147, Essen, Germany.
authors.forename: Kathrin
authors.initials: K
authors.lastname: Kleinehr
authors.affiliation: Department of General-, Visceral-, and Transplantation Surgery, University Hospital of Essen, University of Duisburg-Essen, Hufelandstr. 55, 45147, Essen, Germany.
authors.forename: Gernot M
authors.initials: GM
authors.lastname: Kaiser
authors.affiliation: Department of General-, Visceral-, and Transplantation Surgery, University Hospital of Essen, University of Duisburg-Essen, Hufelandstr. 55, 45147, Essen, Germany; Department of Transplantation Surgery, Semmelweis University, Budapest, Hungary.
authors.forename: Zotlan
authors.initials: Z
authors.lastname: Mathé
authors.affiliation: Department of General-, Visceral-, and Transplantation Surgery, University Hospital of Essen, University of Duisburg-Essen, Hufelandstr. 55, 45147, Essen, Germany.
authors.forename: Juergen-Walter
authors.initials: JW
authors.lastname: Treckmann
authors.affiliation: Department of General-, Visceral-, and Transplantation Surgery, University Hospital of Essen, University of Duisburg-Essen, Hufelandstr. 55, 45147, Essen, Germany.
authors.forename: Andreas
authors.initials: A
authors.lastname: Paul
authors.affiliation: Institute of Virology, University Hospital of Essen, University of Duisburg-Essen, Virchowstr. 179, 45147, Essen, Germany.
authors.forename: Kathrin
authors.initials: K
authors.lastname: Skibbe
authors.affiliation: Institute of Virology, University Hospital of Essen, University of Duisburg-Essen, Virchowstr. 179, 45147, Essen, Germany; Institute of Virology, University Hospital Düsseldorf, Düsseldorf, Germany.
authors.forename: Joerg
authors.initials: J
authors.lastname: Timm
authors.affiliation: Department of Gastroenterology and Hepatology, University Hospital of Essen, University Duisburg-Essen, Hufelandstr. 55, 45147, Essen, Germany.
authors.forename: Ali
authors.initials: A
authors.lastname: Canbay
authors.affiliation: Department of Gastroenterology and Hepatology, University Hospital of Essen, University Duisburg-Essen, Hufelandstr. 55, 45147, Essen, Germany.
authors.forename: Guido
authors.initials: G
authors.lastname: Gerken
authors.affiliation: Department of Gastroenterology and Hepatology, University Hospital of Essen, University Duisburg-Essen, Hufelandstr. 55, 45147, Essen, Germany.
authors.forename: Joerg F
authors.initials: JF
authors.lastname: Schlaak
authors.affiliation: Department of Gastroenterology and Hepatology, University Hospital of Essen, University Duisburg-Essen, Hufelandstr. 55, 45147, Essen, Germany.
authors.forename: Ruth
authors.initials: R
authors.lastname: Broering
mesh_terms: D015415:Biomarkers; D002469:Cell Separation; D042783:Endothelial Cells / Q000166:cytology / Q000378:metabolism; D055166:Hepatic Stellate Cells / Q000166:cytology / Q000378:metabolism; D022781:Hepatocytes / Q000166:cytology / Q000378:metabolism; D006801:Humans; D007728:Kupffer Cells / Q000166:cytology / Q000378:metabolism; D008099:Liver / Q000166:cytology*; D061251:Primary Cell Culture
chemical_list: D015415:Biomarkers