Liver Progenitor Cell Line HepaRG Differentiated in a Bioartificial Liver Effectively Supplies Liver Support to Rats with Acute Liver Failure

Geert A A Nibourg,Ronald P J Oude Elferink,Thomas M Van Gulik,An F C Ruiter,Ruurdtje Hoekstra,Wouter H Lamers,Martinus A W Maas,Robert A F M Chamuleau,Tessa V Van Der Hoeven,Simon Afford

doi:10.1371/journal.pone.0038778

Geert A A Nibourg, Ronald P J Oude Elferink + Show 8 more

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https://doi.org/10.1371/journal.pone.0038778

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Journal: PLoS ONE	Publication Date: Jun 18, 2012
Citations: 74	License type: CC BY 4.0

Affiliation: University of Amsterdam, Academic Medical Center

Abstract

A major roadblock to the application of bioartificial livers is the need for a human liver cell line that displays a high and broad level of hepatic functionality. The human bipotent liver progenitor cell line HepaRG is a promising candidate in this respect, for its potential to differentiate into hepatocytes and bile duct cells. Metabolism and synthesis of HepaRG monolayer cultures is relatively high and their drug metabolism can be enhanced upon treatment with 2% dimethyl sulfoxide (DMSO). However, their potential for bioartificial liver application has not been assessed so far. Therefore, HepaRG cells were cultured in the Academic Medical Center bioartificial liver (AMC-BAL) with and without DMSO and assessed for their hepatic functionality in vitro and in a rat model of acute liver failure. HepaRG-AMC-BALs cultured without DMSO eliminated ammonia and lactate, and produced apolipoprotein A-1 at rates comparable to freshly isolated hepatocytes. Cytochrome P450 3A4 transcript levels and activity were high with 88% and 37%, respectively, of the level of hepatocytes. DMSO treatment of HepaRG-AMC-BALs reduced the cell population and the abovementioned functions drastically. Therefore, solely HepaRG-AMC-BALs cultured without DMSO were tested for efficacy in rats with acute liver failure (n = 6). HepaRG-AMC-BAL treatment increased survival time of acute liver failure rats ∼50% compared to acellular-BAL treatment. Moreover, HepaRG-AMC-BAL treatment decreased the progression of hepatic encephalopathy, kidney failure, and ammonia accumulation. These results demonstrate that the HepaRG-AMC-BAL is a promising bioartificial liver for clinical application.

Highlights

Acute liver failure (ALF) and acute-on-chronic liver failure are severe clinical syndromes with mortality rates as high as 80% [1,2]
In HepaRG-Academic Medical Center bioartificial liver (AMC-Bioartificial livers (BALs)) cultured for an additional 14 days in the presence of 2% dimethyl sulfoxide (DMSO) (+DMSO group), the total DNA content decreased to 25% compared to the pre-BAL and the 2DMSO groups.The total AST and lactate dehydrogenase (LDH) content per BAL were 134668 and 3126100 U, respectively
We demonstrated that the HepaRG-AMC-BAL displays a broad and a high level of hepatic functionality in vitro, as measured by the elimination of ammonia, production of urea, production of 6b-hydroxytestosterone, consumption of lactate, and synthesis of apolipoprotein A-1

Summary

Introduction

Acute liver failure (ALF) and acute-on-chronic liver failure are severe clinical syndromes with mortality rates as high as 80% [1,2]. The syndromes present as a severe impairment of liver function with hepatocellular necrosis, leading to hepatic encephalopathy (HE), systemic inflammation, and multi-organ failure. Despite the progress made in supportive care, liver transplantation is often the only cure, increasing the survival rates to over 80% [3,4]. Liver transplantation is limited by the scarcity of donor organs.In the US, about 20% of the patients with severe liver failure (MELD score .30) who are on the waiting list for liver transplantation die while waiting for a donor liver [5]. Due to xenotransplantation-related risks, there is an urgent need for BAL systems relying on human biocomponents [9]

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