Orthosiphon stamineus Benth. is a medicinal plant belongs to Lamiaceae family. Due to the high variations in rosmarinic acid produced from the fi eld-derived plants, cell suspension culture techniques was previously suggested and practiced as the alternative for cell biomass and rosmarinic acid (RA) production. In the present study, the effects of medium pH, photoperiod, temperature and sucrose were evaluated on cell biomass and rosmarinic acid production using three different cell lines of O. stamineus. This study revealed that the slow and intermediate growing cell lines had similarities in terms of growth pattern. However, cells cultured in medium supplemented with lower sucrose concentration (30 g/L) were more productive in terms of cell biomass and RA in the intermediate and fast growing cell lines however slow growing cells cultured in proliferation medium with 45 g/l sucrose produced higher amount of RA. Medium pH in the range of 5.65-5.8 was optimal for both cell biomass and RA production. Photoperiod did not have a consistent effect on the three cell line's biomass production but the alternation of darkness and illumination led to high RA production in the slow growing cell line. This study proved that abiotic factors including medium pH, photoperiod, temperature and carbohydrate source had impacts on cell biomass and rosmarinic acid production although in a diverse range among different growing cell lines.
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