A study was conducted to elucidate the effects of using different media (MS, ½MS, VW) supplemented with different combinations of 15% coconut water, 10% potato extract and 5% mashed banana with an extra 0.2% activated charcoal on seed germination of Dendrobium signatum Rchb.f. in In vitro cultures. The results showed that the most suitable media, providing 100% seed germination, were MS added to 10% potato extract, ½MS supplemented with 10% potato extract and MS added to 5% mashed banana. Young plantlets were then transferred to MS and ½MS medium supplemented with various kinds of cytokinin including BA, Kinetin and TDZ at the concentrations of 0, 1 and 2 mg/l, and combined with 0 and 0.5 mg/l of NAA for plantlet development. The results showed that young shoots could be differentiated and regenerated into plantlets in all the treatments. The results revealed that the most suitable medium for shoot proliferation and root induction was ½MS medium with 2 mg/l BA added and combined with 0.5 mg/l NAA.