Archives of Toxicology

, Volume 93, Issue 7, pp 1965–1978 | Cite as

Functional transepithelial transport measurements to detect nephrotoxicity in vitro using the RPTEC/TERT1 cell line

  • Philipp F. Secker
  • Nadja Schlichenmaier
  • Mario Beilmann
  • Ulrich Deschl
  • Daniel R. DietrichEmail author
Organ Toxicity and Mechanisms


The kidney is a frequent target for organ-specific toxicity as a result of its primary function in controlling body fluids, for example, via resorption of amino acids, peptides, nutrients, ions, xenobiotics and water from the primary urine as well as excretion of metabolic waste products and hydrophilic and amphiphilic xenobiotics. Compounds exhibiting dose-limiting nephrotoxicity include drugs from highly diverse classes and chemical structures, e.g., antibiotics (gentamicin), chemotherapeutics (cisplatin), immunosuppressants (cyclosporine A and tacrolimus) or bisphosphonates (zoledronate). All of these compounds elicit nephrotoxicity primarily by injuring renal proximal tubule epithelial cells (RPTECs). However, prediction of a compound’s nephrotoxic potential in humans to support early unmasking of risk-bearing drug candidates remains an unmet challenge, mainly due to the complex kidney anatomy as well as pronounced inter- and intraspecies differences and lack of relevant and validated human in vitro models. Accordingly, we used the recently established human RPTEC/TERT1 cell line to carry out toxicity studies with a focus on impairment of functional characteristics, i.e., transepithelial electrical resistance (TEER), vectorial transport of water, cations, and anions. Results were compared to real-time cytotoxicity assessments using cellular impedance (xCELLigence assay) and the routine cell viability readout (MTT). As expected, most toxins caused exposure time- and concentration-dependent cytotoxicity. However, for some compounds (cyclosporine A and tacrolimus), transport processes were strongly impaired in absence of a concomitant decrease in cell viability. In conclusion, these data demonstrate that functional parameters are important, highly sensitive and meaningful additional readouts for nephrotoxicity assessment in human renal proximal tubule epithelial cells.


Kidney Nephrotoxicity Proximal tubule In vitro Epithelial transport 


Supplementary material

204_2019_2469_MOESM1_ESM.docx (15 kb)
Supplementary file1 (DOCX 14 kb)


  1. Aschauer L, Carta G, Vogelsang N, Schlatter E, Jennings P (2015a) Expression of xenobiotic transporters in the human renal proximal tubule cell line RPTEC/TERT1. Toxicol In Vitro 30(1 Pt A):95–105. CrossRefPubMedGoogle Scholar
  2. Aschauer L, Limonciel A, Wilmes A et al (2015b) Application of RPTEC/TERT1 cells for investigation of repeat dose nephrotoxicity: a transcriptomic study. Toxicol In Vitro 30(1 Pt A):106–16. CrossRefGoogle Scholar
  3. Asphahani F, Zhang M (2007) Cellular impedance biosensors for drug screening and toxin detection. Analyst 132(9):835–841. CrossRefPubMedPubMedCentralGoogle Scholar
  4. Duff T, Carter S, Feldman G et al (2002) Transepithelial resistance and inulin permeability as endpoints in in vitro nephrotoxicity testing. Altern Lab Anim 30(Suppl 2):53–59CrossRefGoogle Scholar
  5. Gai Z, Visentin M, Hiller C et al (2016) Organic cation transporter 2 overexpression may confer an increased risk of gentamicin-induced nephrotoxicity. Antimicrob Agents Chemother 60(9):5573–5580. CrossRefPubMedPubMedCentralGoogle Scholar
  6. Hall AM, Unwin RJ (2007) The not so 'mighty chondrion': emergence of renal diseases due to mitochondrial dysfunction. Nephron Physiol 105(1):p1–10. CrossRefPubMedGoogle Scholar
  7. Hausherr V, van Thriel C, Krug A, Leist M, Schobel N (2014) Impairment of glutamate signaling in mouse central nervous system neurons in vitro by tri-ortho-cresyl phosphate at noncytotoxic concentrations. Toxicol Sci 142(1):274–284. CrossRefPubMedGoogle Scholar
  8. Jenkinson SE, Chung GW, van Loon E, Bakar NS, Dalzell AM, Brown CD (2012) The limitations of renal epithelial cell line HK-2 as a model of drug transporter expression and function in the proximal tubule. Pflugers Arch 464(6):601–611. CrossRefPubMedGoogle Scholar
  9. Lepist EI, Ray AS (2016) Renal transporter-mediated drug-drug interactions: are they clinically relevant? J Clin Pharmacol 56(Suppl 7):S73–81. CrossRefPubMedGoogle Scholar
  10. Lin Z, Will Y (2012) Evaluation of drugs with specific organ toxicities in organ-specific cell lines. Toxicol Sci 126(1):114–127. CrossRefPubMedGoogle Scholar
  11. Loboz KK, Shenfield GM (2005) Drug combinations and impaired renal function—the 'triple whammy'. Br J Clin Pharmacol 59(2):239–243. CrossRefPubMedPubMedCentralGoogle Scholar
  12. Masereeuw R, Moons MM, Toomey BH, Russel FG, Miller DS (1999) Active lucifer yellow secretion in renal proximal tubule: evidence for organic anion transport system crossover. J Pharmacol Exp Ther 289(2):1104–1111PubMedGoogle Scholar
  13. Miller RP, Tadagavadi RK, Ramesh G, Reeves WB (2010) Mechanisms of Cisplatin nephrotoxicity. Toxins (Basel) 2(11):2490–2518. CrossRefGoogle Scholar
  14. Morrissey KM, Stocker SL, Wittwer MB, Xu L, Giacomini KM (2013) Renal transporters in drug development. Annu Rev Pharmacol Toxicol 53:503–529. CrossRefPubMedGoogle Scholar
  15. Naesens M, Kuypers DR, Sarwal M (2009) Calcineurin inhibitor nephrotoxicity. Clin J Am Soc Nephrol 4(2):481–508. CrossRefPubMedGoogle Scholar
  16. Paueksakon P, Fogo AB (2017) Drug-induced nephropathies. Histopathology 70(1):94–108. CrossRefPubMedGoogle Scholar
  17. Pazhayattil GS, Shirali AC (2014) Drug-induced impairment of renal function. Int J Nephrol Renovasc Dis 7:457–468. CrossRefPubMedPubMedCentralGoogle Scholar
  18. Perazella MA, Markowitz GS (2008) Bisphosphonate nephrotoxicity. Kidney Int 74(11):1385–1393. CrossRefPubMedGoogle Scholar
  19. Pfaller W, Gstraunthaler G (1998) Nephrotoxicity testing in vitro–what we know and what we need to know. Environ Health Perspect 106(Suppl 2):559–569CrossRefGoogle Scholar
  20. Puri S, Folias AE, Hebrok M (2015) Plasticity and dedifferentiation within the pancreas: development, homeostasis, and disease. Cell Stem Cell 16(1):18–31. CrossRefPubMedGoogle Scholar
  21. Quiros Y, Vicente-Vicente L, Morales AI, Lopez-Novoa JM, Lopez-Hernandez FJ (2011) An integrative overview on the mechanisms underlying the renal tubular cytotoxicity of gentamicin. Toxicol Sci 119(2):245–256. CrossRefPubMedGoogle Scholar
  22. Sauzay C, White-Koning M, Hennebelle I et al (2016) Inhibition of OCT2, MATE1 and MATE2-K as a possible mechanism of drug interaction between pazopanib and cisplatin. Pharmacol Res 110:89–95. CrossRefPubMedGoogle Scholar
  23. Secker PF, Luks L, Schlichenmaier N, Dietrich DR (2018) RPTEC/TERT1 cells form highly differentiated tubules when cultured in a 3D matrix. Altex 35(2):223–234. CrossRefPubMedGoogle Scholar
  24. Selen A, Amidon GL, Welling PG (1982) Pharmacokinetics of probenecid following oral doses to human volunteers. J Pharm Sci 71(11):1238–1242CrossRefGoogle Scholar
  25. Sepand MR, Ghahremani MH, Razavi-Azarkhiavi K et al (2016) Ellagic acid confers protection against gentamicin-induced oxidative damage, mitochondrial dysfunction and apoptosis-related nephrotoxicity. J Pharm Pharmacol 68(9):1222–1232. CrossRefPubMedGoogle Scholar
  26. Sirenko O, Grimm FA, Ryan KR et al (2017) In vitro cardiotoxicity assessment of environmental chemicals using an organotypic human induced pluripotent stem cell-derived model. Toxicol Appl Pharmacol 322:60–74. CrossRefPubMedPubMedCentralGoogle Scholar
  27. Srinivasan B, Kolli AR, Esch MB, Abaci HE, Shuler ML, Hickman JJ (2015) TEER measurement techniques for in vitro barrier model systems. J Lab Autom 20(2):107–126. CrossRefPubMedPubMedCentralGoogle Scholar
  28. Tiong HY, Huang P, Xiong S, Li Y, Vathsala A, Zink D (2014) Drug-induced nephrotoxicity: clinical impact and preclinical in vitro models. Mol Pharm 11(7):1933–1948. CrossRefPubMedGoogle Scholar
  29. Waring MJ, Arrowsmith J, Leach AR et al (2015) An analysis of the attrition of drug candidates from four major pharmaceutical companies. Nat Rev Drug Discov 14(7):475–486. CrossRefPubMedGoogle Scholar
  30. Wenting-Van Wijk MJ, Blankenstein MA, Lafeber FP, Bijlsma JW (1999) Relation of plasma dexamethasone to clinical response. Clin Exp Rheumatol 17(3):305–312PubMedGoogle Scholar
  31. Wieser M, Stadler G, Jennings P et al (2008) hTERT alone immortalizes epithelial cells of renal proximal tubules without changing their functional characteristics. Am J Physiol Renal Physiol 295(5):F1365–F1375. CrossRefPubMedGoogle Scholar
  32. Wilmer MJ, Saleem MA, Masereeuw R et al (2010) Novel conditionally immortalized human proximal tubule cell line expressing functional influx and efflux transporters. Cell Tissue Res 339(2):449–457. CrossRefPubMedGoogle Scholar
  33. Wilmes A, Limonciel A, Aschauer L et al (2013) Application of integrated transcriptomic, proteomic and metabolomic profiling for the delineation of mechanisms of drug induced cell stress. J Proteomics 79:180–194. CrossRefGoogle Scholar
  34. Wilmes A, Aschauer L, Limonciel A, Pfaller W, Jennings P (2014) Evidence for a role of claudin 2 as a proximal tubular stress responsive paracellular water channel. Toxicol Appl Pharmacol 279(2):163–172. CrossRefPubMedGoogle Scholar
  35. Wilmes A, Bielow C, Ranninger C, et al. (2015) Mechanism of cisplatin proximal tubule toxicity revealed by integrating transcriptomics, proteomics, metabolomics and biokinetics. Toxicol In Vitro 30(1 Pt A):117–27. CrossRefPubMedGoogle Scholar
  36. Zsengeller ZK, Ellezian L, Brown D et al (2012) Cisplatin nephrotoxicity involves mitochondrial i njury with impaired tubular mitochondrial enzyme activity. J Histochem Cytochem 60(7):521–529. CrossRefPubMedPubMedCentralGoogle Scholar

Copyright information

© Springer-Verlag GmbH Germany, part of Springer Nature 2019

Authors and Affiliations

  1. 1.Human and Environmental Toxicology, Department of BiologyUniversity of KonstanzKonstanzGermany
  2. 2.Brain Cancer Metabolism GroupGerman Cancer Research Center (DKFZ)HeidelbergGermany
  3. 3.Boehringer Ingelheim Pharma GmbH & Co. KG, Non-Clinical Drug SafetyBiberachGermany

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