TY  - THES
T1  - Organotypic functional cultures of human liver cells for long-term maintenance and assessment of drug-induced metabolome effects
A1  - Müller,Daniel
Y1  - 2012/04/26
N2  - The goals of this thesis were (i) to establish and improve organotypic liver cell culture techniques for long-term pharmacological studies and (ii) to develop and apply a metabolomics based approach for the assessment of drug-induced effects. As first model, a 3D bioreactor system was characterized in terms of cell physiology and functionality. Primary human hepatocytes could be kept viable and functional for more than 2 weeks in this system. Optimization of the system allowed determination of oxygen uptake rates and viability. As second 3D system, a hanging drop method was successfully applied for the generation of organotypic cultures as high-throughput in vitro model for toxicity studies. 
For the investigation of drug-induced metabolic effects, a metabolomics approach based on GCTOF-MS combined with multivariate statistics was developed. In 2D cultures of primary human hepatocytes, both short-term (up to 4 days) and long-term (3 weeks) drug-induced effects were detected at clinically relevant concentrations, showing the sensitivity of the established method. 
Both 3D cultivation methods used in this thesis represent a step forward to organotypic cultures as in vitro alternatives to animals and are particularly suitable for the investigation of chronic toxicity. The established metabolomics approach is a sensitive tool to assess drug-induced changes even at subtoxic concentrations and can be applied to other cell types and cultivation systems in the future.
KW  - Metabolom
KW  - Leberepithelzelle
KW  - Organkultur
KW  - In-vitro-Kultur
CY  - Saarbrücken
PB  - Universitäts- und Landesbibliothek
AD  - Postfach 151141, 66041 Saarbrücken
UR  - http://scidok.sulb.uni-saarland.de/volltexte/2012/4836
ER  -