Malignant peripheral nerve sheath tumor (MPNST) is a rare malignant tumor, for which only a few cultured cell lines are available to date. In the present study, we established three new MPNST cell lines, FU-SFT8611, FU-SFT8710 and FU-SFT9817, from a 40-year-old Japanese man without neurofibromatosis 1 (NF1), a 43-year-old Japanese woman with NF1, and a 61-year-old Japanese woman without NF1, respectively. These cell lines were in culture for more than 3 years in vitro, and exhibit complex karyotypes lacking a common characteristic pattern. Two of the cell lines, FU-SFT8611 and FU-SFT9817, were tumorigenic in nude mice, with the resultant tumors showing an immunohistochemical phenotype similar to the original tumors. Comparative genomic hybridization analysis revealed that chromosomal imbalances were very similar between the original tumors and the established cell lines, but no consistent imbalances among the three cell lines as reported so far. As for chromosomal imbalances that have been reported to be associated with poor survival, FU-SFT8611 exhibited a gain of chromosome 17q22-qter, and FU-SFT9817 showed a loss of 17q22-qter, while gains of 17q24 and 7p15-p21 were observed in none of the cell lines. These newly established cell lines provide a valuable resource for biological and pathological investigations into new treatment regimes for MPNST.