In the present study, to 3-month-old shoots of Paphiopedilum callosum were used as source material for shoot regeneration. 2- and 3-month-old shoots cultured on SH medium supplemented with 1.0 mg/l TDZ, 0.3 mg/l NAA, 30 g/l sucrose and 9 g/l agar gave the best shoot regeneration (4.50 shoots/explant). Explants were efficiently surface sterilized by dipping in 0.1 percent fresh aqueous mercuric chloride (HgCl2) solution for 6 minutes. SH medium supplemented with 0.5 mg/l BA, 0.5 mg/l NAA, 30 g/l sucrose, 9 g/l agar, and 1 g/l activated charcoal was found to be suitable for the shoot growth and development. After 60 days of culture, utilizing the technique of wounding the shoot apex together with application of liquid SH medium in combination with cotton wool supplemented with TDZ at 0.4 - 0.6 mg/l and 30 g/l sucrose resulted in the highest number of shoots (4.50 shoots/explant). Root formation were successfully obtained when these shoots were transferred to SH medium supplemented with 1.0 mg/l NAA, 0.5 mg/l BA, 30 g/l sucrose, 9 g/l agar, and 1 g/l activated charcoal after 60 days of culture. All plantlets with well developed roots (100 percent) survived after the acclimatization period under greenhouse sonditions. This is the first time the technique of wounding the shoot apex was applied in P. callosum, and it showed to be a simple and effective micropropagation method for this recalcitrant species. Moreover, the technique should be considered for the breeding program of other species which belong to the genus Paphiopedilum.