Eight-week old axillary shoots and leaves induced from epicotyls were used as explants for adventitious shoot regeneration in Fraxinus excelsior. The explants were cultured on Murashige and Skoog (MS) or Woody Plant Medium (WPM), supplemented with 2.25 or 4.5 µM Thidiazuron (TDZ) plus 0.49 µM indole-3-butyric acid (IBA) on which callus formation was induced. For all genotypes, satisfactory number of adventitious shoots (2.3 ± 0.3 to 20.3 ± 6.0) was achieved only from axillary shoot callus, cultured on different MS medium, and the duration of cultivation and genotype had the most significant effect on regeneration rate. The rate of survival and regeneration capacity from leaf explants was significantly lower and depended on the genotype. Adventitious shoots with length 2 cm or 3 cm were cultured on half-strength WPM supplemented with 4.9 µM indole-3-butyric acid (IBA) and 5.37 µM α-naphthaleneacetic acid (NAA). After 24 h the shoots were planted in pots, containing peat and perlite (1 : 1, v/v). The shoots were covered with transparent plastic vessels to provide high air humidity and grown in a cultivation chamber. After 4 weeks the cover vessels were removed and all plantlets were transferred to a greenhouse. The longer shoots (3 cm) survived in the cultivation chamber and were acclimatized in greenhouse (75.9 ± 3.0%).