The effect of rat acute-liver-failure plasma on HepaRG cells

Int J Artif Organs. 2012 Nov;35(11):1006-14. doi: 10.5301/ijao.5000121.

Abstract

Purpose: We recently demonstrated the high liver functionality of the human liver cell line HepaRG, including ammonia eliminating capacity, making it a valuable biocomponent of a bioartificial liver (BAL) to support patients with acute liver failure (ALF). This cell line further gains detoxification properties when cultured with dimethyl sulfoxide (DMSO). In this paper we describe whether its functionality is compromised by the toxic effects of ALF plasma, as has been shown for primary hepatocytes.

Methods: We exposed -DMSO and +DMSO HepaRG cultures during 16 hours to healthy plasma and ALF-rat plasma. The cultures were analyzed for lipid accumulation, cell leakage, apolipoprotein A-1 production, nitrogen metabolism and transcript levels of hepatic genes.

Results: The -DMSO cultures showed increased cell leakage after healthy and ALF plasma exposure in contrast to +DMSO cultures, but otherwise the -DMSO and +DMSO cultures were equally affected by exposure to the plasmas. Exposure to both plasmas caused lipid accumulation and decreased transcript levels of various hepatic genes. ALF plasma decreased urea cycle activity, but increased urea production from arginine by upregulated arginase 2. However, total ammonia elimination was not affected by exposure to either plasma, indicating its predominant elimination by fixation into amino acids. In addition, apolipoprotein A-1 production remained constant.

Conclusions: HepaRG cells are negatively affected by rat plasma, even of healthy origin. However, their ammonia eliminating capacity is relatively resistant, underlining their suitability for BAL application. DMSO pre-treatment may increase their viability in plasma.

Publication types

  • Research Support, Non-U.S. Gov't

MeSH terms

  • Animals
  • Arginase / metabolism
  • Cell Culture Techniques
  • Cell Line
  • Cryoprotective Agents / pharmacology
  • Dimethyl Sulfoxide / pharmacology
  • Hepatocytes / drug effects
  • Hepatocytes / physiology*
  • Humans
  • Lipid Metabolism / physiology
  • Liver Failure, Acute / blood*
  • Liver Failure, Acute / pathology*
  • Liver, Artificial
  • Plasma / physiology*
  • Rats

Substances

  • Cryoprotective Agents
  • Arginase
  • Dimethyl Sulfoxide