ATLA::Alternatives to Laboratory Animals

Volume 30, Supplement 2

Transepithelial resistance and inulin permeability as endpoints in in vitro nephrotoxicity testing

ATLA 30, Supplement 2, 53-59, 2002

Tracey Duff,¹ Simon Carter,² Gemma Feldman,³ Gordon Mcewan,¹ Walter Pfaller,⁴ Pauline Rhodes,² Michael Ryan³ and Gabrielle Hawksworth¹

Department of Medicine and Therapeutics and Department of Biomedical Sciences, Polwarth and IMS Building, University of Aberdeen, Foresterhill, Aberdeen A825 2ZD, UK; ²GIaxoSmithOne, Park Road, Ware, Herts, SG12 ODP, UK; ³Department of Pharmacology, University College Dublin, Belfield, Dublin 4, Republic of Ireland; ⁴1nstitute of Physiology, University of Innsbruck, Fritz-Preglstrasse 3, 6010 Innsbruck, Austria

SUMMARY

Transepithelial electrical resistance (RT) and the flux of fluorescein isothiocyanate (FITC) across Madin Darby canine kidney (MDCK) strain 1 cells and porcine epithelial kidney (LLC-PK1) monolayers were compared between three laboratories for a range of nephrotoxins. The precision of the REMS AutoSampler was similar to that of the Ussing chamber and the ENDOHM® technique, but superior to using chopstick electrodes, for measurements of resistance. The nephrotoxins used were selective for the proximal tubule, and in all cases, LLC-PK1 cells were more sensitive than MDCK cells. In most cases, change in RT was a more sensitive indicator of damage than alterations in FITC flux. The REMS system provides high intra-plate precision for RT measurements and is a higher throughput system, which is applicable to screening for nephrotoxicity in vitro.

Keywords: in vitro testing, medium-throughput screening, nephrotoxicity, nephrotoxins, transepithelial resistance, transmembrane flux