ymphytum officinale L. (Boraginaceae) is a well-known medicinal plant and a source of natural compounds with high antioxidant activity. The initiation of “in vitro” cultures of Symphytum officinale L aimed not only to assess the dedifferentiation capacity depending on explant origin and growth regulators, but also to develop a multiplication protocol based on indirect regeneration through shoots, followed by roots development induction. The proliferative capacity was tested on leaf and shoots explants, cultivated on Murashige-Skoog basal medium ( MS ), testing two auxins:: naphtalenacetic acid (NAA) and indolylacetic acid (IAA) and two cytokinines: kinetine (K) and benzylaminopurine (BAP). The MS medium with 1.0 mg/l IAA and 0,1 mg/l BAP proved to be the best for callus induction from leaf explants. Shoot regeneration was achieved after subculturing the calli on MS medium supplemented with 1 mg/l BAP and 0,1 mg/l IAA. It was found to be the best for multiple shoot regeneration from callus through organogenesis Multiple shoot proliferation was noticed at 3th subculture in medium and shoot proliferation was decreased with the increased number of subculture. Root system development was achieved on MS medium without growth regulators. Rooted shoots (plantlets) were gradually acclimatized.