In vitro propagation of Smilax glabra Roxb. was studied. The results indicated that sterilized explants cultured on MS + 6-BA 1.0 mg/L + NAA 0.1 mg/L could lead to budding fastly, the medium MS + 6-BA 1.0 mg/L + NAA 0.1 mg/L + 15% CM suitable for the proliferation, 3/2MS + 6-BA 0.05 mg/L + 4% Sugar suitable for buds strong. Treatment on the medium H (modified) + NAA 0.5 mg/L was best for rooting. The survival rate of the test-tube plantlets was 95% in transplanting on media contained turf-sand (1:1).