In the present study trials were done to micropropagate Hylocereus costaricencis newly introduced to Egypt. Surface sterilized areole's sections (as a starting material) were subjected to various factorial concentrations of benzyladenine, kinetin and thiadiazhuron at 0.5, 1.0 and 1.5 mg/l added to MS culture media to induce shoot morphogenesis. The obtained shoots were rooted using indole butyric acid and/or naphthalene acetic acid individually, combined and with or without addition of activated charcoal. The results obtained revealed that cytokinins could successfully induce multiple shoot (mini-joints) morphogenesis and the response depended on their types and concentrations. After 12 weeks of treatment the number of shoots obtained using BA, Kin and TDZ at 1.5 mg/l reached up 13.67±2.03, 8.67±1.2 and 8.67±1.2 respectively in comparison with 1.0±0.58 shoots per control explant. The maximum number of rootlets 4.00±0.58 per shoot was obtained in response to the combination of IBA/NAA at 0.25mg/l for 12 weeks,. The addition of charcoal (0.1%) reduced the number of rootlets from 4.00±0.58 to 3.00±0.58 with concomitant increase in the root length from 0.83±0.17 to 4.17±0.44. The rooted regenerants were transferred to pots containing a mixture of peat moss: sand: clay in the ratio of 1:1:1 with a 100% survival regardless of the type of treatments used for either shoot or root induction. The obtained results indicate that in vitro micropropagation of H. costaricensis can be used as a modern biotechnological tool to meet with the growing local market demand and help the spread of this newly introduced plant in Egypt.