The successful in vitro cultures establishment was achieved in Rosa × alba ‘Maxima’, Rosa gallica ‘Cardinal de Richelieu’, Rosa helenae ‘Semiplena’, and Rosa × centifolia ‘Paul Ricault’. After surface disinfection with 15% “Domestos”, shoot tips were cultured on Murashige and Skoog (1962) (MS) medium supplemented with 5 µM N⁶-benzyladenine (BA). Regeneration depended on the rose cultivar, and 64-100% of explants developed shoots. The best result in axillary shoot multiplication was observed on MS medium with 5 or 10 µM BA and the highest shoot multiplication rate was obtained in Rosa helenae ‘Semiplena’ both during establishing the culture as well as during the multiplication of axillary shoots. In the rooting experiments half-strength MS medium, supplemented with 2.5 µM indole-3-butyric acid (IBA) and 240 µM phloroglucinol (PG) were tested, and in addition to traditional fluorescence lamps, red light emitting diodes was applied. Red light had no impact on shoot rooting, which was optimal on half-strength MS medium, without IBA and PG.