Aspalathus linearis, commonly known as rooibos, is a commercially important species of the Fabaceae family, endemic to the Fynbos biome in the Cape Floristic region of South Africa. Rooibos has been shown to have strong antioxidant activity, as well as anti-inflammatory, hepatoprotective, antimutagenic and UV-protective activity. As such, rooibos in traditional and holistic healthcare has been used for relieving skin and digestive disorders, mild depression, nervous tension, and insomnia. It is hypothesised that the bioactivity of A. linearis is due to the overall synergistic effect of the metabolite content unique to this species. Amongst the several phenolic flavonoids found in A. linearis, are aspalathin and nothofagin, two key compounds, of which the former is unique to the species. Other flavonoids present, such as vitexin, orientin and rutin have known anticancer activity. Several ecotypes of rooibos exist which differ significantly from the cultivated ecotype with regards to morphology, genotype and chemotype. Previous work has shown that cultivated A. linearis has anticarcinogenic properties, however, no pharmacological studies have yet been performed on the other existing ecotypes. This study, therefore, aimed to identify potential applications of wild-growing populations of A. linearis for the treatment of melanoma using in vitro cell based assays. Five population extracts were tested for their effect on the cell proliferation of the human melanoma cell line, A375, as well as their cytotoxicity on human keratinocyte (HaCaT) cells, using an MTT cell viability assay. Significant decreases in A375 cell proliferation were shown by all five extracts, with two populations exhibiting higher activity in the 72-h dose response trial with IC50 values of below 100 µg/ml. While the cytotoxicity values of these ecotypes on healthy skin cells are relatively low in comparison, rendering these extracts as promising additions to existing melanoma treatments, such as immunotherapy.