Nephrotoxicity pp 725-730 | Cite as

Isolated Rat Renal Proximal Tubular Cells: A Model to Investigate Drug Induced Nephrotoxicity

  • E. M. Gordon
  • P. H. Whiting
  • J. G. Simpson
  • G. M. Hawksworth


The kidney shows marked functional, morphological and biochemical heterogeneity which may account for the site-specific toxicity of several drugs and xenobiotics. For example, the toxicity associated with cephaloridine and some aminoglycosides is confined to the proximal tubular cells (Kuo and Hook, 1982; Kaloyanides and Pastoriza-Munoz, 1980), whereas prolonged intake of paracetamol results in renal papillary necrosis (Mohandas et al., 1984). To investigate the mechanisms of toxicity using cell suspensions or primary cultures, it is therefore necessary to isolate the different cell types. Several approaches have been used to obtain preparations of proximal tubular cells or fragments. One approach uses existing cell lines of kidney origin (LLC-PK1 or MDCK), the disadvantage being that the exact site of origin within the nephron is not known and, being a cell line, it may not be totally representative of the normal physiological state. A second approach is the digestion or explantation of kidney tissue in the presence of serum-free, hormonally defined culture media such that fibroblast proliferation is minimal and epithelial cell growth is encouraged (Chuman et al., 1982). Unfortunately both these cell preparations lack a brush border, which may be critical for the active uptake of drugs and chemicals.


Proximal Tubular Cell Amino Acid Precursor Define Culture Medium Hank Solution Cytochrome P450 Content 


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  1. Allen, M. B., Blair, J.McD., 1972, The regulation of rabbit liver fructose-1, 6-diphosphatase activity by phospholipids in vivo, Biochem. J., 130:1167PubMedGoogle Scholar
  2. Bach, P.H., Ketley, C. P., Ahmed, I., Dixit, M., 1986, The mechanisms of target cell injury by nephrotoxins, Fd. Chem. Toxic., 24: 775CrossRefGoogle Scholar
  3. Burke, M.D., Mayer, R.T., 1974, Ethoxyresoruf in: direct fluorimetric assay of a microsomal O-dealkylation which is preferentially inducible by 3-methylcholanthrene, Drug Metab. Dispos., 2: 583PubMedGoogle Scholar
  4. Burke, M.D., Thompson, S., Elcombe, C. R., Halpert, J., Haaparanta, T., Mayer, R.T., 1985, Ethoxy-, pentoxy- and benzyloxy-phenoxazones and 6 homologues. A series of substrates to distinguish between different induced cytochromes P450, Biochem. Pharm., 34:3337PubMedCrossRefGoogle Scholar
  5. Chuman, L., Fine, L.G., Cohen, A.H., Saier, M.H.Jr., 1982, Continuous growth of proximal tubular kidney epithelial cells in hormone-supplemented serum-free medium, J. Cell Biol., 94:506PubMedCrossRefGoogle Scholar
  6. Endou, H., Koseki, C., Haumara, S., Kakuno, K., Hojo, K., Sakai, F., 1982, Renal cytochrome P450; its localisation along a nephron and its induction, In: “Biochemistry of Kidney Functions”, F. Morel, ed., Elsevier Biomedical PressGoogle Scholar
  7. Gordon, E.M., McDougall, S.M., Whiting, P.H., Hawksworth, G.M., 1987, Gentamicin-stimulated increase in PAH uptake in renal slices and isolated proximal tubular cells, this volume.Google Scholar
  8. Gordon, E.M., Whiting, P.H., Simpson, J.G., Hawksworth, G.M., 1987, Isolation and characterisation of rat renal proximal tubular cells, Biochem. Soc. Trans., 15:457Google Scholar
  9. Goujon, F.M., Nebert, D.W., Gielen, J.E., 1972, Genetic expression of aryl hydrocarbon hydroxylase induction, Mol. Pharm., 8:667Google Scholar
  10. Guder, W. G., Ross, B.D., 1984, Enzyme distribution along the nephron, Kid. Int., 26:101CrossRefGoogle Scholar
  11. Habig, W.H., Jakoby, W.B., 1981, Assays for determination of glutathione S-transferases, Meth. Enzymol., 77:398PubMedCrossRefGoogle Scholar
  12. Hissin, P.J., Hilf, R., 1976, A fluorimetric method for determination of oxidised and reduced glutathione in tissues, Analvt. Biochem., 74:214CrossRefGoogle Scholar
  13. Jones, D.P., Sundby, G., Ormstad, K., Orrenius, S., 1979, Use of isolated kidney cells for study of drug metabolism, Biochem. Pharm., 28:929PubMedCrossRefGoogle Scholar
  14. Kaloyanides, G.J. and Pastoriza-Munoz, E., 1980, Aminoglycoside nephrotoxicity, Kid. Int., 18:571CrossRefGoogle Scholar
  15. Kuo, C., Hook, J.B., 1982, Depletion of renal glutathione content and nephrotoxicity of cephaloridine in rabbits, rats and mice, Toxic Appl. Pharm., 63:292CrossRefGoogle Scholar
  16. Kuo, C., Maita, K., Sleight, S.D., Hook, J.B., 1983, Lipid peroxidation: A possible mechanism of cephaloridine-induced nephrotoxicity, Toxic. Appl. Pharm., 67:78CrossRefGoogle Scholar
  17. Mohandas, J., Marshall, J. J., Duggin, G. G., Horvarth, J. S., Tiller, D.J., 1984, Differential distribution of glutathione and glutathione related enzymes in rabbit kidney: Possible implications in analgesic nephropathy, Biochem. Pharm., 33:1801PubMedCrossRefGoogle Scholar
  18. Orrenius, S., Ellin, A., Jakobson, S.V., Thor, H., Cinti, D. L., Schenkman, J.B., Estabrook, R.W., 1973, The cytochrome P450-containing mono-oxygenase system of rat kidney cortex microsomes, Drug Metab. Dispos., 1:350PubMedGoogle Scholar
  19. Rosenberg, M.R., Michalopoulos, G., 1987, Kidney proximal tubular cells isolated by collagenase perfusion grow in defined media in the absence of growth factors, J. Cell Physiol., 131:107PubMedCrossRefGoogle Scholar
  20. Schmidt, U., Marosvari, I., Dubach, U.C., 1975, Renal metabolism of glucose: Anatomical sites of hexokinase activity in the rat nephron, FEBS Letts., 53:26CrossRefGoogle Scholar
  21. Smith, M.A., Acosta, D., Bruckner, J.V., 1986, Development of a primary culture system of rat kidney cortical cells to evaluate the nephrotoxicity of xenobiotics, Fd. Chem. Toxic., 24:551CrossRefGoogle Scholar
  22. Smith, M.A., Acosta, D., Bruckner, J.V., 1987, Cephaloridine toxicity in primary cultures of rat renal cortical epithelial cells, Toxicol. In Vitro, 1:23PubMedCrossRefGoogle Scholar
  23. Vinay, P., Gougoux, A., Lemieux, G., 1981, Isolation of a pure suspension of rat proximal tubules, Am. J. Physiol., 241:403Google Scholar

Copyright information

© Springer Science+Business Media New York 1989

Authors and Affiliations

  • E. M. Gordon
    • 1
    • 2
    • 3
  • P. H. Whiting
    • 2
  • J. G. Simpson
    • 4
  • G. M. Hawksworth
    • 2
    • 3
  1. 1.Clinical Pharmacology UnitUniversity of AberdeenForesterhill, AberdeenUK
  2. 2.Department of Chemical PathologyUniversity of AberdeenForesterhill, AberdeenUK
  3. 3.Department of PharmacologyUniversity of AberdeenForesterhill, AberdeenUK
  4. 4.Department of PathologyUniversity of AberdeenForesterhill, AberdeenUK

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