The present study was undertaken to compare the alkaloid composition of Ochrosia elliptica callus tissue with that of in vivo plants, as no previous studies had examined the biosynthetic capabilities of its cultured tissues. Calli were initiated from stem, root, and leaf explants of young seedlings on modified Gamborg B5 or Linsmaier and Skoog media. Alkaloids were isolated via Soxhlet extraction, purified by preparative thin-layer chromatography (TLC), and identified using spectral (UV, fluorimetry, mass spectrometry) and physicochemical methods by comparison with standards. Results confirmed the presence of ellipticine and 9-methoxyellipticine in callus tissues, with their levels being almost equivalent to those in in vivo leaves. Traces of alkaloids were also detected in the culture medium. The fluorometric method developed was effective for quantifying these alkaloids. Studies are ongoing to produce these antitumor alkaloids in a bioreactor.