Tanacetum parthenium has gained attention for drug development due to its production of bioactive secondary metabolites, such as phenolic compounds (PHEC) and parthenolide (PTN). This work investigated the dierences in the production of those compounds among in vitro cultures (organs from plantlets and roots cultures) of T. parthenium. Results showed that the in vitro cultures produced PHEC and PTN, which depended on the type of culture and the incubation time. The content of secondary metabolites was higher in shoots of plantlets (S-ivPL), followed by root cultures (R-ivC) and roots of plantlets (R-ivPL). The PHEC and PTN production was enhanced in plantlets by applying a combination of plant growth regulators (PGRs: -naphtalenacetic acid [NAA] at 0.27 M with kinetin [KIN] at 2.32 M). The highest content of secondary metabolites (62.54 mg gallic acid equivalents g1; 4.261 and 4.973 mg chlorogenic acid g1; 0.858 and 1.015 mg caeic acid g1; 1.931 mg salicylic acid g1; 0.071 mg PTN g1) was found in the PGRs-treated S-ivPL samples. Also, morphogenesis and growth of plantlets was enhanced by these PGRs.