Melanoma Cell Culture Protocol

The assessment was performed on radial growth phase-WM35 human melanoma (Wistar Institute, Philadelphia, PA, USA) [20 (link)] cells and a highly metastatic melanoma cell line M1-15 [21 (link)], donated by professor Andras Falus, Genetics Department, University Semmelweis, Budapest. Melanoma cells were cultured in RPMI medium supplemented with 5% fetal calf serum, 50 μg/ml gentamicin and 5 ng/ml amphotericin, all from Biochrom AG (Berlin, Germany), to avoid the medium influence on the cell lines properties, as previously described [22 (link), 23 (link)]. For all experiments, the cells were used within 4 passages, to preserve their original melanoma characteristics [24 (link), 25 (link)]. Cultures were fed twice weekly and incubated in a humid atmosphere at 37°C and 5% CO₂. All experiments were conducted in triplicate in subdued light, as previously described [26 (link)].