Three independent experiments were carried out in order to establish criteria for the process of acclimatization of henequen plants and their pursuit in the nersery stage. Plants were micropropagated following the protocol developed by González (2001). In the first experiment the size effect of plant let size on survival rate during acclimatization was evaluated. Best answers were obtained for calibers higher than 6.9 cm. In a second experiment the effect of different levels of organic matter (OM) in the substratum was evaluated in the sustrato. Best results are obtained supplementing substratum with 10% of organic matter. The third experiment analyses the effect of different substrate and plantation parameters in nursery state. The results showed that the condition of the substratum plays a significant role in acclimatization of micropropagated plants of henequen.