Objective: To study the effects of physical and chemical factors on hairy root of Panax ginseng and its glycoside production. Methods: The cell growth index and its glycoside yield in different culture condition such as different plant hormones mixed, mediums and time were compared. Results: After hairy root of Panax ginseng was precultivated on the MS medium with IBA (0.5%) at first 72 hours. the culture was cultured on the MS medium. During 6 weeks, dry weight of hairy root increased by 98.74 times, and the content of glycoside synthesized by hairy root is 5.190% and Re+Rg1 is 0.3271%, respectively. Conclusion:We have achieved the best condition of hairy root of Panax ginseng.