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Cell Biology and Toxicology

, Volume 12, Issue 4–6, pp 289–298 | Cite as

Precision-cut dog renal cortical slices in dynamic organ culture for the study of cisplatin nephrotoxicity

  • H. J. Toutain
  • J. P. Sarsat
  • A. Bouant
  • D. Hoet
  • D. Leroy
  • V. Moronvalle-Halley
Plenary lectures Culture of renal cells

Abstract

The dog is the non-rodent species the most often used in preclinical drug safety evaluation. In this study, we established a new system of precision-cut dog renal cortical slices, evaluated their biochemical, functional, and morphological integrity, and determined the effects of cisplatin (cis-diamminedichloroplatinum (II), CDDP), a very potent nephrotoxic antineoplastic agent used to treat a variety of solid tumors, on the viability and histopathology of slices. Precision-cut renal cortical slices were made perpendicular to the cortical-papillary axis. Slices were incubated in DMEM/Ham's F12 culture medium containing 1 g/L glucose, 2 mmol/L glutamine, and 2 mmol/L heptanoic acid at 37°C in an atmosphere of 5% CO2-70% O2-25% N2 in dynamic organ culture. Our results showed that slices maintained ATP and GSH content, protein synthesis, Na+-dependent uptake of glucose inhibited by phlorizin, PAH (p-aminohippuric acid) uptake inhibited by probenecid, and TEA (tetraethylammonium) uptake inhibited by mepiperphenidol for at least 6 h of culture, and morphological integrity up to 24 h. After 6 h of exposure, CDDP induced vacuolation and cell necrosis in the epithelial tubular cells of slices with a concentration-related increase in extension but not in severity. The development of the lesions started in the proximal tubules and extended to the distal tubules. The location and the extension of the lesions confirmed the observations in dog kidneys after in vivo treatment with CDDP by the intravenous route. The concentration-related decrease in slice viability after 6 h exposure to CDDP was in keeping with the extension of the histopathological lesions in the renal parenchyma. The slice viability was unaffected up to 0.63 mmol/L CDDP. At 1.25 and 2.5 mmol/L CDDP, slice viability fell by 35% and 75%, respectively. These results suggest that precision-cut dog renal cortical slices in culture may be suitable for addressing the specific nephrotoxicity issues encountered in this species.

Keywords

renal cortical slices dog hispathology glutathione transport systems cisplatin nephrotoxicity 

Abbreviations

CDDP

cis-diamminedichloroplatinum (II)

FIS

freshly isolated slices

GSH

glutathione

GSSG

glutathione disulfide

Hepes

4-(2-hydroxyethyl)-1-piperazineethanesulfonic acid

MGP

methyl-α-D-glucopyranoside

MTT

3-[4,5-dimethylthiazol-2-yl]-2,5-diphenyltetrazolium bromide

PAH

p-aminohippuric acid

PBS

phosphate-buffered saline

TEA

tetraethylammonium

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Copyright information

© Kluwer Academic Publishers 1996

Authors and Affiliations

  • H. J. Toutain
    • 1
  • J. P. Sarsat
    • 1
  • A. Bouant
    • 1
  • D. Hoet
    • 1
  • D. Leroy
    • 1
  • V. Moronvalle-Halley
    • 1
  1. 1.Service de Toxicologie ExpérimentaleDépartment Sécurité du MédicamentVitry sur SeineFrance

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