Ruscus aculeatus L. (Liliaceae, Butcher’s broom) is a small evergreen shrub used both in gardening and as cut foliage. The plant’s rhizomes are widely collected as a drug source of steroid saponins with antiinflammatory, venotonic and antihaemorrhoidal activity. The species is under regulated regime of gathering in Bulgaria. The natural reproduction of R. aculeatus is very slow and despite of the limited resources its cultivation is not widespread. Seeds and whole plants were collected from natural populations in Southeast Bulgaria. Preliminary experiments showed that rhizomes and cladodes were unsuitable as primary explants. In vitro shoot cultures were initiated from mature embryos on MS media, supplemented with 2 mg l^-1 NAA for two months. The effect of NAA and BAP on regeneration capacity of rhizome explants was tested. Best shoot proliferation was observed in rhizome explants by consequent cultivation in liquid and agar-solidified media. The obtained regenerants rooted spontaneously and were directly transferred to greenhouse conditions. In vitro micropropagation of R. aculeatus could be an effective way for conservation and cultivation of the species.