Abstract: In order to acquire the suspension cultivation production that cells growth quickly, biomass liveweight larger and edysterone content higher, the inducing condition for obtaining the granular embryonic callus was studied by chosen culture medium and screened the phytohormone, and the Ajuga lobata D. Don cell suspension culture system was established by optimizing from three aspects, which were the culture medium type, plant hormone species and their proportion, amount of inoculation in this paper. The results shown that: MS solid medium added 0.4 mg.L-1 2,4-D could induce the Ajuga lobata D.Don’s root segments to form unconsolidated granular embryogenic callus. Comparing the growth status of cell lines in three kind of liquid culture medium such as WPM, MS, 1/2 MS found that cells were grew best in MS. 2,4-D affected cell growth obviously. The cell biomass was 1.3 times higher than which cultured in culture medium added NAA. Experiments on the effect of phytohormone ratio on the Ajuga .lobata D. Don cell suspension culture shown that ,with 6-BA concentration increasing, ecdysterone content increased. Compared with that in control group, edysterone content in the cells cultured in medium added 0.5mg.L- 6–BA was significantly higher (P<0.05). The cells cultured in liquid medium added 1mg.L- 6-BA accumulated very significantly higher edysterone than that accumulated in the cells in control group (P<0.01). Though, the most prosper phytohormone proportion was 0.4 mg.L 1 2,4-D+6-0.5mg.L 1 BA，under which the net cell biomass could reach 4.3652±1.0739g after cultured 7 days, the ecdysterone content could reach 4.5692±0.2044mg/g of cell dry weight ,the cell lines was more uniform and grew faster. When the inoculation amount was 15%, cell lines grew best, at the 7th day after inoculation, cell growth rate was 1.109g/d,cell regeneration index can reach 3.45. It was the most proper inoculation quantity. Conclusion: the optimal cell suspension culture system was established , which would benefit the ecdysterone production.