The characteristics of ferric nitrilotriacetate (Fe-NTA)-induced rat renal cell carcinoma by intraperitoneal administration and the effects of hormones on the tumor have been studied in vivo. However, to our knowledge there have been no reports on the characteristics of the tumor cells. In this study, we established a tumor cell line from Fe-NTA induced rat renal tumour and investigated tumor cell characteristics. The tumor cell line RKFe was established from Fe-NTA induced rat renal tumor by collagenase digestion. RKFe showed epithelial morphology in culture, and had microvilli and basement membranes under electron microscopy. RKFe cells acquired tolerance to cytotoxicity of Fe-NTA. They also grew in soft agar culture and formed tumors by the inoculation of cells into nude mice. Histologically, transplanted tumors showed poorly defferentiated adenocarcinoma with several tubular structures. A subclone of RKFe, named RKFe-C6, showed metastasis to lungs. The RKFe cell line may be useful for studying the characteristics of Fe-NTA induced rat tumor cells.