title: Induction of an Inflammatory Response in Primary Hepatocyte Cultures from Mice.
abstract: The liver plays a decisive role in the regulation of systemic inflammation. In chronic kidney disease in particular, the liver reacts in response to the uremic milieu, oxidative stress, endotoxemia and the decreased clearance of circulating proinflammatory cytokines by producing a large number of acute-phase reactants. Experimental tools to study inflammation and the underlying role of hepatocytes are crucial to understand the regulation and contribution of hepatic cytokines to a systemic acute phase response and a prolonged pro-inflammatory scenario, especially in an intricate setting such as chronic kidney disease. Since studying complex mechanisms of inflammation in vivo remains challenging, resource-intensive and usually requires the usage of transgenic animals, primary isolated hepatocytes provide a robust tool to gain mechanistic insights into the hepatic acute-phase response. Since this in vitro technique features moderate costs, high reproducibility and common technical knowledge, primary isolated hepatocytes can also be easily used as a screening approach. Here, we describe an enzymatic-based method to isolate primary murine hepatocytes, and we describe the assessment of an inflammatory response in these cells using ELISA and quantitative real-time PCR.
authors.affiliation: Katz Family Drug Discovery Center and Division of Nephrology and Hypertension, Department of Medicine, University of Miami Leonard M. Miller School of Medicine; Department of Cell Biology and Anatomy, University of Miami Leonard M. Miller School of Medicine.
authors.forename: Brian
authors.initials: B
authors.lastname: Czaya
authors.affiliation: Katz Family Drug Discovery Center and Division of Nephrology and Hypertension, Department of Medicine, University of Miami Leonard M. Miller School of Medicine; Department of Cell Biology and Anatomy, University of Miami Leonard M. Miller School of Medicine.
authors.forename: Saurav
authors.initials: S
authors.lastname: Singh
authors.affiliation: Katz Family Drug Discovery Center and Division of Nephrology and Hypertension, Department of Medicine, University of Miami Leonard M. Miller School of Medicine; Department of Cell Biology and Anatomy, University of Miami Leonard M. Miller School of Medicine.
authors.forename: Christopher
authors.initials: C
authors.lastname: Yanucil
authors.affiliation: Katz Family Drug Discovery Center and Division of Nephrology and Hypertension, Department of Medicine, University of Miami Leonard M. Miller School of Medicine; Department of Cell Biology and Anatomy, University of Miami Leonard M. Miller School of Medicine.
authors.forename: Karla
authors.initials: K
authors.lastname: Schramm
authors.affiliation: Katz Family Drug Discovery Center and Division of Nephrology and Hypertension, Department of Medicine, University of Miami Leonard M. Miller School of Medicine; Department of Cell Biology and Anatomy, University of Miami Leonard M. Miller School of Medicine.
authors.forename: Christian
authors.initials: C
authors.lastname: Faul
authors.affiliation: Katz Family Drug Discovery Center and Division of Nephrology and Hypertension, Department of Medicine, University of Miami Leonard M. Miller School of Medicine; alexander.grabner@duke.edu.
authors.forename: Alexander
authors.initials: A
authors.lastname: Grabner
mesh_terms: D000818:Animals; D018929:Cell Culture Techniques / Q000379:methods; D002478:Cells, Cultured; D016207:Cytokines / Q000378:metabolism; D004797:Enzyme-Linked Immunosorbent Assay / Q000379:methods; D022781:Hepatocytes / Q000166:cytology*; D007249:Inflammation / Q000139:chemically induced / Q000378:metabolism / Q000473:pathology*; D018836:Inflammation Mediators / Q000378:metabolism; D015850:Interleukin-6 / Q000009:adverse effects; D008070:Lipopolysaccharides / Q000633:toxicity; D008099:Liver / Q000166:cytology*; D051379:Mice; D010477:Perfusion; D060888:Real-Time Polymerase Chain Reaction / Q000379:methods*; D015203:Reproducibility of Results
chemical_list: D016207:Cytokines; D018836:Inflammation Mediators; D015850:Interleukin-6; D008070:Lipopolysaccharides