Characterization of a Rat Multi-Cell Type 3D-Liver Microtissue System
- Corresponding Author:
- Alex Odermatt
Division of Molecular and Systems Toxicology
Department of Pharmaceutical Sciences, University of Basel
Klingelbergstrasse 50, 4056 Basel, Switzerland
Tel: +41 61 267 1530
Fax: +41 61 267 1515
E-mail: [email protected]
Received date: April 08, 2013; Accepted date: November 20, 2013; Published date: November 22, 2013
Citation: Kratschmar DV, Messner S, Moritz W, Odermatt A (2013) Characterization of a Rat Multi-Cell Type 3D-Liver Microtissue System. J Tissue Sci Eng 4:130. doi:10.4172/2157-7552.1000130
Copyright: © 2013 Kratschmar DV, et al. This is an open-access article distributed under the terms of the Creative Commons Attribution License, which permits unrestricted use, distribution, and reproduction in any medium, provided the original author and source are credited.
Abstract Background: Primary hepatocytes rapidly lose their polarized morphology and the expression of important liverspecific metabolic enzymes, receptors and transport proteins under normal two-dimensional (2D) culture conditions. Thus, their use as a reliable predictive in vitro model for drug-induced liver injury is limited. Three-dimensional (3D) liver micro tissue culture systems have become an increasingly attractive alternative for the evaluation of druginduced liver toxicity. However, several liver-specific pathways remain to be characterized in such models. Methods and principal findings: In the present work, we compared the expression of several genes with a role in the anti-oxidant cell defense and glucocorticoid pathways in rat H4IIE hepatoma cells, primary rat hepatocytes, 2D-hepatocyte sandwich culture, and a multi-cell type micro tissue model comprising primary rat hepatocytes in coculture with liver-derived nonparenchymal cells and macrophage (Kupffer cells). Gene expression was studied for up to 25 days in culture. High expression levels of the Nrf2-dependent genes NQO1, ABCC3 and GST2A were detected in H4IIE cells and in 3D-liver microtissues, in contrast to 2D-hepatocytes where a rapid decline of these genes was observed. The glucocorticoid-dependent genes ORM1, G6PC, PCK1 and HSD11B1 were highly expressed up to 25 days of cultivation in 3D-liver microtissues, but they showed very low or background expression in H4IIE and 2D-hepatocyte models. Conclusions: The tested 3D-multi-cell type liver micro tissue represents a stable and functionally active model system, with sustained expression for more than three weeks of cultivation of important metabolic proteins regulated by the glucocorticoid and anti-oxidant cell defense pathway.