Assessment of a new cell culture perfusion apparatus for in vitro chronic toxicity testing. Part 1: technical description.
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.
Koppelstaetter
Christian
C
Department of Physiology, University of Innsbruck, A-Innsbruck, Austria.
Jennings
Paul
P
Ryan
Michael P
MP
Morin
Jean-Paul
JP
Hartung
Thomas
T
Pfaller
Walter
W
D000826:Animal Testing Alternatives; D000818:Animals; D018929:Cell Culture Techniques / Q000295:instrumentation / Q000379:methods*; D004396:Coloring Agents; D003470:Culture Media; D006801:Humans; D006863:Hydrogen-Ion Concentration; D007674:Kidney Diseases / Q000139:chemically induced; D007687:Kidney Tubules, Proximal / Q000166:cytology / Q000378:metabolism*; D007770:L-Lactate Dehydrogenase / Q000378:metabolism; D010078:Oxazines; D010477:Perfusion / Q000295:instrumentation / Q000379:methods; D015203:Reproducibility of Results; D013696:Temperature; D013997:Time Factors; D036401:Toxicity Tests, Chronic / Q000295:instrumentation* / Q000379:methods*; D014966:Xanthenes
D004396:Coloring Agents; D003470:Culture Media; D010078:Oxazines; D014966:Xanthenes; C005843:resazurin; D007770:L-Lactate Dehydrogenase