Micropropagation is a technique of rapid in vitro clonal propagation of plants, which enables their high frequency multiplication. Adventitious regeneration is a process of in vitro production of new plant tissues and organs from non-meristematic cells. The aims of the master thesis were to develop protocols for sterilization of cannabis seeds, establishment of aseptic cultures, micropropagation and adventitious regeneration from different explants (hypocotyl and cotyledon), to test the usefulness of ISSR markers for detection of somaclonal variation and to check our hypotheses. In the experiments, we used already known plant tissue culture media and we tested some new media formulations for axillary shoots and roots initiation of plants. The best medium for the formation of the axillary shoots was MS basal medium supplemented with the hormone cytokinine TDZ and the longest shoots were initiated from the third and fourth nodes on the plant, while the apical shoots were less suitable for initiation of an in vitro shoot culture. MS basal medium supplemented with hormone cytokinine meta-topolin proved to be most suitable for rooting. In vitro regeneration of hemp is genotype dependent and the cotyledon explants proved to be more regenerative than hypocotyl explants. Our results with ISSR markers showed that they are useful for genetic differentiation between different genotypes of hemp.