Review Article

Advances in predictive in vitro models of drug-induced nephrotoxicity

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In vitro screens for nephrotoxicity are currently poorly predictive of toxicity in humans. Although the functional proteins that are expressed by nephron tubules and mediate drug susceptibility are well known, current in vitro cellular models poorly replicate both the morphology and the function of kidney tubules and therefore fail to demonstrate injury responses to drugs that would be nephrotoxic in vivo. Advances in protocols to enable the directed differentiation of pluripotent stem cells into multiple renal cell types and the development of microfluidic and 3D culture systems have opened a range of potential new platforms for evaluating drug nephrotoxicity. Many of the new in vitro culture systems have been characterized by the expression and function of transporters, enzymes, and other functional proteins that are expressed by the kidney and have been implicated in drug-induced renal injury. In vitro platforms that express these proteins and exhibit molecular biomarkers that have been used as readouts of injury demonstrate improved functional maturity compared with static 2D cultures and represent an opportunity to model injury to renal cell types that have hitherto received little attention. As nephrotoxicity screening platforms become more physiologically relevant, they will facilitate the development of safer drugs and improved clinical management of nephrotoxicants.

Key points

  • Currently available in vitro and animal models of drug-induced nephrotoxicity are poorly predictive of toxicity in humans.

  • Functional proteins that underlie the susceptibility of various renal cell types to specific drugs, and molecular biomarkers of injury, can be used to characterize the functional maturity of in vitro models and their capacity to respond to nephrotoxicants.

  • In vitro models derived using new protocols for the directed differentiation of pluripotent stem cells to renal cells and new 3D in vitro culture systems demonstrate improved functional maturity over static 2D systems.

  • Improved functional maturity of cultured renal cells in systems that more closely replicate the physiology of the renal tubule and its supporting cells will improve the predictive ability of in vitro models of nephrotoxicity.

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J.Y.-C.S. is supported by a University of Melbourne Research Scholarship and a Murdoch Children’s Research Institute Top up scholarship. J.J. is supported by the Dutch Kidney Foundation (Kolff postdoctoral fellowship abroad grant 170KK05), EMBO (short-term fellowship 6893), and by the partners of Regenerative Medicine Crossing Borders powered by Health~Holland, Top Sector Life Sciences & Health. R.M. is supported by a grant from the UK National Centre for the Replacement, Refinement and Reduction of Animals in Research (NC3Rs), the NephroTube CRACK IT Challenge, and the RegMed XB consortium. M.H.L. is an Australian National Health and Medical Research Council (NHMRC) Senior Principal Research Fellow (GNT1042093) and is supported by funding from the NHMRC (GNT1100970) and the US National Institutes of Health (DK107344).

Reviewer information

Nature Reviews Nephrology thanks J. Lewis, R. Morizane and the other, anonymous reviewer(s) for their contribution to the peer review of this work.

Author information

Author notes

  1. These authors contributed equally: Joanne Y-C Soo, Jitske Jansen


  1. Department of Paediatrics, The University of Melbourne, Parkville, Victoria, Australia

    • Joanne Y.-C. Soo
    •  & Melissa H. Little
  2. Murdoch Children’s Research Institute, Parkville, Victoria, Australia

    • Joanne Y.-C. Soo
    •  & Melissa H. Little
  3. Division of Pharmacology, Utrecht Institute for Pharmaceutical Sciences, Utrecht University, Utrecht, Netherlands

    • Jitske Jansen
    •  & Rosalinde Masereeuw
  4. Department of Anatomy and Neuroscience, The University of Melbourne, Parkville, Victoria, Australia

    • Melissa H. Little


  1. Search for Joanne Y.-C. Soo in:

  2. Search for Jitske Jansen in:

  3. Search for Rosalinde Masereeuw in:

  4. Search for Melissa H. Little in:


J.J. researched data for the article. J.Y.-C.S., J.J., and R.M. wrote the article. All authors contributed substantially to discussion of the article’s content and reviewed and edited the manuscript before submission.

Competing interests

M.H.L. holds a research contract with Organovo, Inc. The other authors declare no competing interests.

Corresponding author

Correspondence to Melissa H. Little.



Any compounds, natural and synthetic, that exert an adverse effect on a specific kidney cell type or mediate an unwanted event affecting kidney functioning. By contrast, a toxin is a poisonous substance produced within living cells or organisms.

Electrogenic transport

Transport that leads to a change in net charge across a cell membrane.

Transepithelial electrical resistance

(TEER). The electrical resistance across a cell monolayer. The higher the value, the less permeable the monolayer.


Renal cells seeded in a 2D or 3D configuration in a microfluidic device. For proximal tubule chips, these devices typically allow flow of media across the cells’ apical surface, basolateral surface, or both. Other cell types such as endothelial cells might be included; in these cases, the organization of the different cell types is defined by the design of the chip.

Kidney organoids

Three-dimensional aggregates of interstitial cells and nephron structures with characteristic segments, typically formed by directing pluripotent stem cells to a renal fate and aggregating these cells to enable self-organization, with or without additional extracellular matrix.

Renal tissue arrays

Three-dimensional co-cultures of renal epithelial cells, renal fibroblasts, and endothelial cells. Cell suspensions are prepared in biocompatible gels and bioprinted. The composition of the suspensions and the spatial arrangement of the different suspensions used define the organization of the different cell types. Scaffolds composed of extracellular matrix may or may not be used.

Fugitive ink

Biocompatible material that can be printed and later evacuated to leave a hollow space within a mould.

Biofunctionalized hollow fibres

Hollow, porous polymer fibres coated with extracellular matrix on which renal cells can be cultured.


Peeling and shedding of the top layer of an epithelium.