Aconitine alkaloids produced from cell suspension cultures of Aconitum napellus for the first time. The effects of various culture conditions on cell biomass and aconitine accumulation in cell suspension cultures were investigated. Suspension cell cultures of A. napellus were established by transferring callus tissues from leaf explants onto liquid MS medium supplemented with 1 mg/l NAA and 0.1 mg/l kinetin. Among the culture media tested, MS medium had a pronounced effect on cell growth and aconitine accumulation. The maximum dry cell weight was obtained at inoculum size of 3 g (FCW) per flask and in MS medium supplemented with 5% sucrose after 8 weeks. The addition of salicylic acid (SA) and yeast extract (YE) in the MS medium enhanced aconitine accumulation. Using a proper combination of culture condition and supplements, aconitine content could reach 0.043% (dry weight basis), that was 2.5~3 fold higher that detected in control cultures.