A new human cell line (Wa-2) derived from an extrarenal rhabdoid tumor has been established. The cell line grows as a monolayer consisting of round- and spindle-shaped cells. Injection of cells into nude mice results in the growth of solid tumors within 2 wk of inoculation. These solid tumors have the microscopic appearance similar to that of the original tumor from which the cell line was derived. Moreover, the tumor cells have all the features of rhabdoid tumors, including the intracytoplasmatic hyaline globules, large prominent nucleoli, and inclusion bodies made up of whorls of fibrillary material. Transplanted tumor cells stain positive for vimentin, cytokeratin, actin, and desmin and negative for myoglobin and neuron-specific enolase. Karyotyping of the cell line at different passages and cells derived from the transplant tumors consistently revealed a diploid number of chromosomes with a reciprocal translocation between chromosomes 18 and 22 [t(18;22) (q21;p11.2)]. In fibroblasts derived from the patient, no translocation could be found. Culturing the cells in the presence of 1-beta-D-arabinofuranosylcytosine induces differentiation, characterized by the outgrowth of neuron-like processes and the morphological appearance of cells similar to neuroblasts. Southern blot analysis showed no amplification of the N-myc oncogene. Since no published cell line derived from rhabdoid tumors exists, this cell line should be helpful to further elucidate the biology and histological origin of the malignant rhabdoid tumor.