The experiment was conducted at the period from 20th Dec. 2015 to 30th Mar. 2016 to know the effect of growth medium and nodes number on the acclimatization success of tissue propagated potatoes seedlings (Burrin class) and study some plant physiological properties. The first experiment involved testing growth media: loam, peat moss, and perlite with different portions (1:1) to plant tissue propagated potato tubers, while the second experiment involved testing the nodes number where 9, 8, 7, 6, and 5 plant nodes were chosen. Results of the first experiment showed the superiority of the culture media of peat moss + perlite to other culture media by giving the higher values of shoot system studied properties: plant height, branches number, leaves number, fresh weight, and dry weight of 15 cm, 2.33 branches, 14.66 leaves, 0.840 g, 0.082 g, respectively, and root system: root length, tubers number, fresh weight, and dry weight of 13.33 cm, 14.33 roots, 3.66 tubers, 0.660 g, and 0.068 g, respectively. The results of second experiment showed the superiority of the 9 nodes to other nodes number by giving the higher values of the studied properties (in the first experiment) for the shoot system of 17.33 cm, 4.33 branches, 15.33 leaves, 0.860 g, and 0.078 g, respectively, while for the root system of 12.33 cm, 8.00 roots, 3.66 tubers, 0.610 g, and 0.072 g, respectively. A conclusion can be drawn that the best growth medium to acclimate potato seedlings was loam + perlite and the 9 nodes was the best number due to their significant superiority compared with other