Culturing Human Melanoma and Control Cell Lines

The human melanoma M285, M375 and M296, cell lines were a kind gift from Antoni Ribas (University of California, Los Angeles), and the M202, A375, M229, SKmel28 and SKmel5 cells were a kind gift from Randall T. Moon and Andy J. Chien (University of Washington, Seattle). Primary human melanocytes adult (HEMa-LP) were obtained from Gibco. Human embryonic kidney cells (HEK-293) and rhabdomyosarcoma cells (RD-1) were obtained from the Biomedical Research Centre (University of East Anglia, UK). Human melanoma cells were cultured as previously described [29 (link)]. HEMa-LP cells were cultured in Medium-254 (Gibco) with the addition of PMA-Free Human Melanocyte Growth Supplement-2 (HMGS-2; Gibco). HEK-293 cells were cultured in Dulbecco’s modified Eagle medium (DMEM) + GlutMAX (Gibco) supplemented with 10% FBS, 1% L-glutamine and penicillin and streptomycin. RD-1 cells were cultured in Dulbecco’s modified Eagle medium (DMEM) + GlutMAX (Gibco) supplemented with 10% FBS and penicillin and streptomycin. All cells were maintained at 37° C in a 5% CO₂ air-humidified incubator, were routinely screened for mycoplasma and not cultured beyond passage 25.