Kalanchoe blossfeldiana Poelln. is an ornamental specie used as a plant for pots and garden. Although are found studies on its propagation by biotechnological methods, the conditions to guarantee high survival rates in the acclimatization stage of in vitro plants have not been described. The aim of this work was to evaluate the development of K. blossfeldianain vitro plants during the acclimatization stage. Rooted plants were used in a culture medium without growth regulators. For planting, two mixtures of substrate were used in proportion 80:20, earthworm humus:zeolite and earthworm humus: sand. During the acclimatization the percentage of survival was calculated at 7, 15 and 30 days, and the plants height was measured at 30 days. In the rooting stage an average of 9.8 roots and a 0.9 cm height per plant was obtained. The survival in the acclimatization was 97.5% for both substrates, and there were no significant differences for the variable height of the plant. According to the results, with the substrates used for the acclimatization of K. blossfeldiana in vitro plants, high percentages of survival can be obtained under the conditions studied in the present work. Keywords:Crassulaceae, earthworm humus, substrae, zeolite