We have preliminary evidence of a stem cell in melanoma among cells expressing the CD133 surface marker. These cells also highly express cancer/testis (CT) antigens, making active cancer immunotherapy a highly attractive therapeutic modality for targeting melanoma stem cells. A major limitation of work on cancer stem cells is access to appropriate cancer tissues and cell lines. Ideally experiments must be performed on tumour specimens freshly excised from patients, but this is impractical for most cancers. We have established a method for culturing melanoma under conditions that have been shown in other cancer types to maintain the phenotype and genotype of the excised tumour within the cancer cell line. In this project novel melanoma cell lines will be characterised using our established methods to optimise their culture, to better qualify their unique properties, and most especially to validate the most effective immune targets for melanoma stem cells.