Here, we found that EVs derived from the culture media of metastatic melanoma cells cultivated at pH 6.5 (“acidified” EVs) demonstrate a higher content of the factors involved in melanoma progression (miR-221, pro-migratory protein SNAI1, stemness marker CD133, and mRNAs coding EGFR, VEGF-α, α3- and V-integrins, and HSP60) than EVs derived from the culture media of the same cells cultivated at pH 7.4 (“normal” EVs). This evidence concerns the gene PROM1 and melanoma.