Epigenetically stabilized primary hepatocyte cultures: a potential sensitive screening tool for non-genotoxic carcinogenicity

Replacing, reducing and refining the use of animals in xenobiotic toxicity testing is being gradually implemented in the EU legislation. Therefore, research in this field and development of alternative methods are becoming more important. As the liver and hepatocytes in particular, are a main target for toxicity and carcinogenicity in the organism, a lot of attention is paid to the establishment of liver-based in vitro models. Primary hepatocytes are the golden standard, but their cultures are prone to progressive dedifferentiation, thereby restricting their use to short-term purposes. In this chapter, we present a method to stabilize the differentiated phenotype of hepatocytes in primary culture by remodelling the chromatin structure by using the histone deacetylase inhibitor trichostatin A. After describing the set up of stabilized primary hepatocyte cultures and providing a troubleshooting guide, some results are shown with respect to their potential applicability for testing of non-genotoxic hepatocarcinogenic substances.