Induction of an inflammatory response in primary hepatocyte cultures from mice

Brian Czaya, Saurav Singh, Christopher Yanucil, Karla Schramm, Christian H Faul, Alexander Grabner

Research output: Contribution to journalArticle

2 Scopus citations

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.

Original languageEnglish (US)
Article numbere55319
JournalJournal of Visualized Experiments
Volume2017
Issue number121
DOIs
StatePublished - Mar 10 2017

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Keywords

  • Chronic kidney disease
  • CRP
  • FGF23
  • Hepatocytes
  • Immunology
  • Inflammation
  • Interleukin-6
  • Issue 121
  • Liver
  • LPS
  • Primary cultured cells

ASJC Scopus subject areas

  • Neuroscience(all)
  • Chemical Engineering(all)
  • Immunology and Microbiology(all)
  • Biochemistry, Genetics and Molecular Biology(all)

Cite this

Czaya, B., Singh, S., Yanucil, C., Schramm, K., Faul, C. H., & Grabner, A. (2017). Induction of an inflammatory response in primary hepatocyte cultures from mice. Journal of Visualized Experiments, 2017(121), [e55319]. https://doi.org/10.3791/55319