This study aimed to investigate the phenolic content, antioxidant activity, cytotoxicity and the in vitro wound healing activity of methanolic and aqueous extracts of Verbascum stachydifolium Boiss & Heldr. var. stachydifolium. Total phenolic and flavonoid contents and antioxidant activity were measured using spectrophotometry-based methods. Quantitative analysis of the selected phenolics was performed by HPLC. The cytotoxic effects of the extracts on L929 mouse fibroblast cells were evaluated by the MTT assay. The migration of treated fibroblast cells was assessed by the cell scratch assay. The expressions of type I collagen, FGF7, TGF-?1 and VEGF were evaluated by qRT-PCR and ELISA. The HPLC-based analysis revealed the presence of different phenolic compounds at varying amounts and high antioxidant activities were detected. The cytotoxicity assay results indicated that the methanolic and aqueous extracts did not exhibit any cytotoxic effect on fibroblast cells when used up to 500 ?g/mL concentration. Fibroblast migration was stimulated to the highest degree by the aqueous extract obtained by maceration as observed in the scratch assay at 60.4% closure. The molecular mechanism of the wound healing activity involves the upregulation of the analyzed genes.