The effective technologies of in vitro establishment and micropropagation of three cultivated strawberry genotypes (Fragaria × ananassa Duch.) acclimatizated for Siberian region (‘Sunny Glade’, ‘June Smile’ and the selected hybrid of 5-90-21) have been developed using the stolon apical meristems as explants. The sterilization of explants was optimized by application of 0.2 % silver nitrate solution as a sterilizing agent. The effects of various 6-benzylaminopurine (BAP) concentrations (0.25–2.0 mg/l) on in vitro explant regeneration potential on Murashige–Skoog (MS) and Gamborg–Eveleigh (B5) media were studied. The best medium variant to initiate the organogenesis from ‘June Smile’ cultivar was MC supplemented with 1.0 mg/l BAP, whereas for 5-90-21 hybrid and ‘Sunny Glade’ cultivar it was B5 with the same BAP concentration. Subsequent cultivation of the developed microshoots on media for micropropagation has shown the effectiveness of B5 medium supplemented with 0.75 mg/l BAP for all genotypes: maximum number of axillary microshoots per explant was amounted to 4.90 ± 0.35 for hybrid 5-90-21, 7.16 ± 0.33 – for ‘Sunny Glade’ and 5.93 ± 0.39 – for ‘June Smile’. The obtained results can be used for development of production system of a healthy planting strawberry material through biotechnological approaches.Key words: cultivated strawberry, micropropagation, Murashige–Skoog´s medium, Gamborg–Eveleigh´s medium, 6-benzylaminopurine.