Protocols in In Vitro Hepatocyte Research

Shinde, Vaibhav, Stöber, Regina, Nemade, Harshal, Sotiriadou, Isaia, Hescheler, Jürgen, Hengstler, Jan, Sachinidis, Agapios, Vaibhav Shinde, Regina Stöber, Harshal Nemade, Isaia Sotiriadou, Jürgen Hescheler, Jan Hengstler, Agapios Sachinidis

Transcriptomics is a powerful tool for high-throughput gene expression profiling. Transcriptome microarray experiments conducted with RNA isolated from hepatocytes after exposure to toxicants enable a deep insight into the molecular mechanisms of hepatotoxicity. This understanding, along with structure-activity relationships underlying hepatotoxicity, will provide a novel strategy to design cost-effective and safer therapeutics. Transcriptomics studies conducted with established hepatotoxic drugs in various in vitro and in vivo hepatotoxicity test systems have contributed to the elucidation of the mechanistic basis of liver insults, which were later on substantiated at the proteomics and metabolomics levels. The present chapter is focused on comprehensive transcriptomics of cultured primary hepatocytes treated with chemicals by applying Affymetrix microarray technology. It also describes the detailed protocol for culturing of hepatocytes, their exposure to toxicants as well as sample collection, including RNA isolation, RNA target preparation and finally the hybridization to gene chips for microarray expression analysis.