Production of valuable secondary metabolites using plant cell, tissue and organ cultures generally hampered by low productivity and residual growth regulators might affect on the safety of products and the health of users. Hairy roots derived from in vitro culture in combination with gene transfer techniques could resolve those problems. The aim of this study was to investigate effect of medium components including mineral composition, sugar content, substrate, pH, temperature, and concentration of activated charcoal on growth of Panax vietnamensis hairy roots infected with Agrobacterium rhizogenes A TCC 15834. After 2 months, the results indicated that SH medium supplemented with 8 gIl agar, 50 gIl sucrose, and pH adjusted to 5.8 - 6.1 was suitable for the growth of hairy root lines. The hairy root lines cultured in the darkness at 25°C resulted in the highest number of roots, root fresh weight and dry weight. Furthennore, adding 1 percent (w/v) activated charcoal into the culture medium led to a positive effect on root proliferation. In addition, hairy root lines cultured in shake flask system was also significantly better than those cultured on semi-solid medium with agar. In this initial study, some hairy root lines of Panax vietnamensis were collected based on different morphological characteristics. These lines would be the good resources for bioreactor culture as well as studies on root biomass of Panax Vietnamensis.