ALTEX :: Alternatives to Animal Experiments

2004, VOLUME 2

Assessment of a New Cell Culture Perfusion Apparatus for In Vitro Chronic Toxicity Testing Part 1: Technical Description

Christian Koppelstaetter1, Paul Jennings2, Michael P. Ryan2, Jean-Paul Morin3, Thomas Hartung4 and Walter Pfaller1

1Department of Physiology, University of Innsbruck, A-Innsbruck; 2Pharmacology Dept., The Conway Institute of Biomolecular and Biomedical Research, IRL-Dublin; 3INSERM, Faculté de Médecine Pharmacie, Université de Rouen, F-Rouen; 4Biomedical Pharmacology, University of Konstanz, D-Germany

SUMMARY

In vitro models for chronic toxicity, defined as a recurring exposure to compounds over a prolonged period of time, are still underrepresented in drug evaluation processes. The classical approach to cell culture is not readily suitable to long term repetitive applications. Therefore, we assessed the use of a commercially available perfusion cell culture apparatus in its applicability to chronic renal toxicity testing and describe the technical aspects of adopting the perfusion cell culture system to our purposes. It was apparent that there is a subtle dynamic difference between human renal proximal tubular cells cultured under perfusion and static conditions as illustrated by the accumulation of lactate dehydrogenase (LDH) and the secondary metabolism of resazurin to hydroresorufin, which occurred only under static conditions. The major achievement was the standardisation of the handling of this system with regard to cell cultivation, pH regulation, temperature regulation, and reproducibility of common toxicity endpoints.

Keywords: perfusion, chronic, toxicity, cell culture