Ovaries were removed from Ostrinia furnacalis (Guenée) pupae and were placed in a flask containing TNM-FH medium with 10% inactivated fetal bovine serum. Cell migration occurred after about 1 wk of the initiation in June 2011. The migrated cells were distributed over most of the flask and were treated with N-methyl-N'-nitro-N-nitrosoguanidine (MNNG), a chemical carcinogen, after about 1 mo of initiation for 26 d. Cells were first subcultured successfully 12 d after the MNNG was removed, followed by subculturing for 30 passages. The established cell line, designated IOZCAS-Osfu-1, were analyzed by DNA fingerprinting-PCR (DAF-PCR) to confirm that it originated from O. furnacalis.