The grapevine, a culture of exceptional importance, expanded in all parts of the world, has been the subject of research in vitro for many years to solve the problems that viticulture encounters in practice. For each individual genotype, a method is sought that would accelerate multiplication under controlled conditions. Cultivation of culture in in vitro conditions means cultivation in glass, in other words, it implies cultivation of plants under controlled conditions under constant temperature and humidity on nutrient media. The composition of the nutrient medium significantly affects the growth and development of the cultivated culture. This work is based on the research of the influence of certain nutrient media on the growth of microcuttings of grapevine varieties: Chardonnay, Graševina, Grk and Pošip, and two genotypes created by crossing two varieties: GRP‐33 and DRP‐7. Varieties differ from each other in the growth rate of microcuttings in in vitro conditions, and the MS nutrient medium proved to be more suitable for growth than the CP nutrient medium. Analysis of the results revealed a significant difference in the height of microcuttings between the genotypes GRP‐33, DRP‐7 and Chardonnay in the first experiment, as well as between the genotypes Grk, Pošip and Graševina in the second experiment. By comparing the values of the second parameter, the number of nodes, there are no significant differences in any of the monitored groups of genotypes.