Murashige and Skoog medium (MS) and modified Anderson's Rhododendron medium (mAN) were compared for in vitro shoot multiplication of three highbush blueberries ‘Berkeley’, ‘Bluecrop’ and ‘Goldtraube’. All media contained 0.5 mg l−1 zeatin applied either alone or combined with 0.1, 1 and 5 mg l−1 IBA. In vitro rooting was induced using mAN medium supplemented with 0.8 mg l−1 IBA and 4 g l−1 activated charcoal. The results obtained showed that mAN medium is more suitable for in vitro multiplication of the selected highbush blueberry cultivars than MS medium. Low concentration of IBA (≤1 mg l−1) added in zeatin-supplemented mAN medium increases shoot multiplication efficiency of highbush blueberries in vitro and can be recommended for large-scale propagation of high-quality plants. MS medium induced partial or full necrosis of stems and leaves, which was more pronounced on media containing zeatin combined with increasing concentration of IBA. Rooting capacity of shoots varied widely among the tested blueberry cultivars. The highest rooting and acclimatization rates were achieved in ‘Goldtraube’ (82.8% and 91.8% respectively), and the lowest (10% and 66.7% respectively) were in ‘Berkeley’.