The Madagascar periwinkle (Catharanthus roseus) produces numerous indole alkaloids, several of which have an important pharmaceutical uses such as ajmalicine. The relationship between hypoxia and ajmalicine production in a cell medium culture of Catharanthus roseus were investigated during the cycle of cell culture, as well as the growth in fresh and dry matter. The results show that the lack of oxygenation in C20D cells provokes a very strong inhibition in accumulation of the alkaloids and of other possible substances. This can be explained by the absence of some downstream enzymes involved in the biosynthesis chain of alkaloids or by the blockage of the upstream terpenes to the production stage. Reoxygenation did not restore hypoxia effect on the alkaloid accumulation. Moreover, the present study showed that the addition of the loganin in the 4th day, in the cell culture medium subjected to hypoxia restored the alkaloid production. Also, the results show that the BAP increases the ajmalicine production. The effect of hypoxia was not shown in the presence of cytokinine. The BAP can without doubt decrease the effects of the hypoxia and increases the ajmalicine production.