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3D Organotypic Liver Cultures and their Application in Predictive Toxicology

3D cultivation allows organotypic microtissue formation ensuring cell-cell contacts and contacts to the extracellular matrix. The communication between cells at tight junctions and across extracellular matrix induces and maintains cellular differentiation, functionality and viability. 3D cell cultivation techniques can tremendously improve studies on drug metabolism, drug toxicity especially long-term repeated dose effects. Using a high-throughput 3D cultivation system, we produced 3D organotypic cultures of human hepatic cells using a 96 well plate based hanging drop method. This method allows scaffold-free reorganization of cells under the force of gravity. We show that the production of 3D organotypic cultures of various hepatic cell lines and primary liver cells is possible (Mueller et al., 2011, 2013). The hepatic cells in these 3D cultures were analyzed for viability and functionality. We show that the organotypic cultures maintain high liver-specific function over 3 weeks of culture (Gunness et al., 2013). The effects of several compounds (acetaminophen, aflatoxin B, valproic acid, chlorpromazine, troglitazone and rosiglitazone) were monitored and compared. Currently we are using this system for mechanistic studies. In conclusion, better functionality of 3D systems improves the prediction of toxicity and will have impact in the changed paradigm of drug screening.

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