The HepaRG cell line is suitable for bioartificial liver application

For bioartificial liver application, cells should meet the following minimal requirements: ammonia elimination, drug metabolism and blood protein synthesis. Here we explore the suitability of HepaRG cells, a human cell line reported to differentiate into hepatocyte clusters and surrounding biliary epithelial-like cells at high density and after exposure to dimethyl sulfoxide (DMSO). The effect of carbamoyl-glutamate (CG), an activator of urea cycle enzyme carbamoylphosphate synthetase (CPS) was studied additionally.The effects of DMSO and/or CG were assessed in presence of 15NH4Cl on HepaRG cells in monolayer. We tested hepatocyte-specific functions at transcript and biochemical level, cell damage parameters and performed immunostainings.Ureagenesis, ammonia/galactose elimination and albumin, glutamine synthetase and CPS transcript levels were higher in −DMSO than +DMSO cultures, probably due to a higher cell content and/or cluster-neighbouring regions contributing to their functionality. DMSO treatment increased cytochrome P450 (CYP) transcript levels and CYP3A4 activity, but also cell damage and repressed hepatic functionality in cluster-neighbouring regions. The levels of ammonia elimination, apolipoprotein A-1 production, and transcription of CYP3A4, CYP2B6 and albumin reached those of primary hepatocytes in either the + or −DMSO cultures. Preconditioning with CG increased conversion of 15NH4Cl into 15N-urea 4-fold only in −DMSO cultures.Hence, HepaRG cells show high metabolic and synthetic functionality in the absence of DMSO, however, their drug metabolism is only high in the presence of DMSO. An unparalleled broad hepatic functionality, suitable for bioartificial liver application, can be accomplished by combining CG treated −DMSO cultures with +DMSO cultures.

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• Cell line HepaRG was tested in monolayer for its potential in bioartificial livers.
• Metabolic and synthetic functionality functions were high in the absence of DMSO.
• DMSO increased drug metabolism, but reduced cell number dramatically.
• Preincubation with carbamoylglutamate increased urea production from ammonia.
• A unique functionality can be accomplished by combining −DMSO with +DMSO cultures.