The kidney forms a frequent target for xenobiotic toxicity. The complex biochemical mechanisms underlying nephrotoxicity are best studied in vitro provided that reliable and relevant in vitro models are available. Since most nephrotoxicants affect primarily the cells of the proximal tubules (PTC), much effort has been directed towards the development of in vitro models of PTC. This review focuses on the preparation of PTC and the use of these cells. Discussed are important criteria such as the viability (survival time) of the cells and the parameters to assess toxicity. Recent studies have shown that isolated PTC in suspension are especially suitable for studies on the biochemical mechanisms of 'acute' nephrotoxicity, whereas PTC in primary culture may be used to investigate mechanisms of nephrotoxic damage at very low concentrations, upon prolonged exposure. PTC cultured on porous filter membranes provide new possibilities to study toxicity in relation to cell and transport polarity. Primary cell cultures of human PTC have been set up. Although a further characterization of these systems is needed, recent data indicate their usefulness.