A promising protocol for enhancement of anthocyanin production in vitro in different cultures of  'Selva', 'Oso grande' and 'Chandler' strawberry cultivars was established. The influence of different concentrations of 2,4‑D and BAP addition to MS‑medium (Murashige and Skoog, 1962) on callus production from shoot tip, leaf and root explants of different aseptic cultures of strawberry cultivars were investigated. The maximum value of growth dynamic of callus fresh weight (gm), dry weight (mg) and dry matter content (%) was recorded with Selva cultivar as compared with other strawberry cultivars. The optimum supplementation for callus production to MS‑medium was 2.0 mg/1  2,4‑D + 0.2 mg/1 BAP. Also, in this respect shoot tips of different strawberry cultivars gave better  results of callus induction than other explants. Meanwhile, the optimum value of total anthocyanin accumulation percentage was recorded with leaf callus cultures of different strawberry cultivars. It was 157.98  μg/gm callus, 134.32 μg/gm callus and 98.73 μg/gm callus in Selva, Chandler and Oso grande, respectively.