The aclimatizacion stage is the critical step during the development of tissue cultures in crops. It was evaluated the response of vitro-plants from pineapple Ananas comosus (L.) Merr., to the applications of the hormones: AlA, AIB and ANA in dos es 0-500-1000 and 1500 ppm at the medium: yellow river sand, natural soil and charcoal-scoria. The number of roots, root length and dry weigh root were recorded in the vitro-plants. In general, the yellow river sand was the medium that showed the best results. The natural soil and the charcoal - scoria had low response without a clear tendency. The AIB hormone presented a very good behaviour with the combination yellow river sand at 1000 and 1500 ppm dose.