Patient Ma-Mel-48 presented with melanoma stage II in December 2000 at the age of 85 years. Progression to stages III and IV was diagnosed in October 2001 and June 2002, respectively. The patient was treated with temozolomide at the beginning of 2003 but never received immunotherapy. In September 2003, the patient died. Patient Ma-Mel-100 presented with melanoma stage II in January 1998 at the age of 72 years. Progression to stage III was diagnosed in September 2002. From February 1998 until September 2002, the patient received adjuvant IFNα therapy. In September 2005, the patient died.
Samples including tumor tissues and peripheral blood mononuclear cells (PBMC) were collected after approval by the institutional review board and patient informed written consent. Tissues were mechanically divided for cryopreservation and generation of the corresponding cell lines. Small tissue pieces were distributed in cell culture dishes, and outgrowing cells were split for the first time at 90% cell confluence. Melanoma cell lines were cultured in RPMI-1640 medium supplemented with glutamine, 10% FCS, and penicillin/streptomycin. Cells were cultured at 37°C in a 5% CO2 atmosphere. Cell lines were authenticated by genetic profiling at the Institute for Forensic Medicine (University Hospital Essen) using the AmpFLSTR-Profiler Plus Kit (Applied Biosystems) and routinely tested every 6 months.
Samples including tumor tissues and peripheral blood mononuclear cells (PBMC) were collected after approval by the institutional review board and patient informed written consent. Tissues were mechanically divided for cryopreservation and generation of the corresponding cell lines. Small tissue pieces were distributed in cell culture dishes, and outgrowing cells were split for the first time at 90% cell confluence. Melanoma cell lines were cultured in RPMI-1640 medium supplemented with glutamine, 10% FCS, and penicillin/streptomycin. Cells were cultured at 37°C in a 5% CO2 atmosphere. Cell lines were authenticated by genetic profiling at the Institute for Forensic Medicine (University Hospital Essen) using the AmpFLSTR-Profiler Plus Kit (Applied Biosystems) and routinely tested every 6 months.