The aim of this study is to improve the propagation of Impatiens balsamina, L by an efficient protocol for in vitro propagation was successfully established through tissue culture technique. Cotyledonary node with sections of cotyledon, section of cotyledons and hypocotyls were derived from 21 days in vitro germinating seeds after surface sterilization. They were cultured for shoot multiplication and callus induction on MS medium fortified with various concentrations of BAP, TDZ or combination between BAP and NAA. It was determined that cotyledonary node with sections of cotyledon on MS medium supplemented with 4.5 mg/L BAP for maximum shoots number (4.33shoots) and callus induction 3.78. The best shoot length (3.08 cm) within 8 weeks from regeneration was achieved on MS medium fortified with 1.0 mg/L BAP + 0.5 mg/L NAA. The number of shoots formed per explants increased by re-culturing for six weeks on the same medium (4.5 mg/L BAP alone), since it produced (9 shoots). While the best shoot length (4cm) was obtained by sub culturing the explants on MS medium received 1.0 mg/L BAP + 0.5 mg/L NAA to MS without growth regulators after six weeks. The best callus fresh weight (9.32 g) and callus volume (7.76 cm³) were recorded with using TDZ at 3.0 mg/L that sub cultured on medium fortified with 4.5 mg/L BAP. In addition the highest dry weight (0.81 g) was obtained by re - culturing on the same medium that contained with 4.5 mg/L BAP.