The research reveals results of the creation of sterile cultures and micropropagation of the species of three unique genera of Cactaceae: Astrophytum asterias (Zucc.) Lem.(І CITES); Blossfeldia liliputiana Werderm.(І І CITES); Strombocactus disciformis (DC.) Britton & Rose.(І CITES). The solutions for sterilization were selected, the time of sterilization, and the sterilization procedure of seeds was approbated. The expedience of usage of stems of juvenile plants grown in vitro for propagation of these species of cacti was proved. The possibility of an obtaining of the necessary quantity of plant material by the way of indirect organogenesis is shown. The proper content of nutrient media and favourable conditions of the cultivation for all stages of the micropropagation were approved.