International Journal of Biological Sciences

Impact factor

ISSN 1449-2288

News feeds of IJBS published articles
Manuscript login

open access Global reach, higher impact

Journal of Genomics - Submit manuscript now...


International Journal of Medical Sciences

Journal of Cancer

Journal of Genomics



Journal of Biomedicine

PubMed Central Indexed in Journal Impact Factor

Int J Biol Sci 2016; 12(8):964-978. doi:10.7150/ijbs.15165

Research Paper

Selecting Cells for Bioartificial Liver Devices and the Importance of a 3D Culture Environment: A Functional Comparison between the HepaRG and C3A Cell Lines

Martien van Wenum1,2, Aziza A.A. Adam1, Theodorus B.M. Hakvoort2, Erik J. Hendriks1, Valery Shevchenko3, Thomas M. van Gulik1, Robert A.F.M. Chamuleau2, Ruurdtje Hoekstra1,2✉

1. Surgical laboratory, Academic Medical Center, University of Amsterdam, the Netherlands.
2. Tytgat Institute for Liver and Intestinal Research, Academic Medical Centre, University of Amsterdam, the Netherlands.
3. Biopredic International, Rennes, France.


Recently, the first clinical trials on Bioartificial Livers (BALs) loaded with a proliferative human hepatocyte cell source have started. There are two cell lines that are currently in an advanced state of BAL development; HepaRG and HepG2/C3A. In this study we aimed to compare both cell lines on applicability in BALs and to identify possible strategies for further improvement. We tested both cell lines in monolayer- and BAL cultures on growth characteristics, hepatic differentiation, nitrogen-, carbohydrate-, amino acid- and xenobiotic metabolism. Interestingly, both cell lines adapted the hepatocyte phenotype more closely when cultured in BALs; e.g. monolayer cultures produced lactate, while BAL cultures showed diminished lactate production (C3A) or conversion to elimination (HepaRG), and urea cycle activity increased upon BAL culturing in both cell lines. HepaRG-BALs outperformed C3A-BALs on xenobiotic metabolism, ammonia elimination and lactate elimination, while protein synthesis was comparable. In BAL cultures of both cell lines ammonia elimination correlated positively with glutamine production and glutamate consumption, suggesting ammonia elimination was mainly driven by the balance between glutaminase and glutamine synthetase activity. Both cell lines lacked significant urea cycle activity and both required multiple culture weeks before reaching optimal differentiation in BALs.

In conclusion, culturing in BALs enhanced hepatic functionality of both cell lines and from these, the HepaRG cells are the most promising proliferative cell source for BAL application.

Keywords: HepaRG, C3A, BAL, Liver support, Hepatocytes, Bioartificial liver.

This is an open access article distributed under the terms of the Creative Commons Attribution (CC BY-NC) License. See for full terms and conditions.
How to cite this article:
van Wenum M, Adam AAA, Hakvoort TBM, Hendriks EJ, Shevchenko V, van Gulik TM, Chamuleau RAFM, Hoekstra R. Selecting Cells for Bioartificial Liver Devices and the Importance of a 3D Culture Environment: A Functional Comparison between the HepaRG and C3A Cell Lines. Int J Biol Sci 2016; 12(8):964-978. doi:10.7150/ijbs.15165. Available from