Cynanchumwilfordii is a traditional Chinese herb and has various pharmacological effects. However, its raw plant material is insufficient to meet the market demand. Adventitious roots (ARs) are a novel alternative material for producing plant products. Herein, C. wilfordii ARs were cultured in 5-L bioreactors to produce bioactive compounds and the effects of salt strength and sucrose concentration were investigated in this study. Furthermore, the kinetic study was implemented for confirming a suitable culture period. Finally, the culture efficiency of different size bioreactors was compared. The full salt strength of Murashige and Skoog medium supplemented with 30 g L−1 sucrose was most favorable for AR biomass and the production of flavonoids, ρ-hydroxyacetophenone, 2,5-dihydroxyacetophenone, and total acetophenone derivatives, whereas the media with 50 and 70 g L−1 sucrose were optimal for the production of 2,4-dihydroxyacetophenone and total polysaccharides, respectively. The accumulation of AR biomass and three acetophenone derivatives peaked on day 25, but those of total polysaccharide and flavonoids peaked on day 30, indicating that the selection of culture period should depend on the target compound to be produced. Among 5-, 10-, and 20 L-bioreactors, the 20-L bioreactor exhibited good culture efficiency. The findings suggest that bioactive compounds of C. wilfordii can be mass produced through AR culture in an established bioreactor system, and the produced ARs can be used as promising raw material in the production of plant products.