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Stevia rebaudiana is beneficial to treat diabetes because of its low-calorie glucoside sweeteners. Natural and vegetative propagation are inefficient. In vitro techniques are an attractive alternative but low propagation and reproducibility rates have been reported. Therefore, different ways to increase natural sweetener production in vitro, such as BIT^®, are required. We compared semisolid medium, liquid medium and BIT^® in terms of Stevia biomass and steviol glycosides production. At 21 days of culture, morphological quality of BIT^®-derived shoots was best and coupled with shoot fresh and dry weight that were more than seven times higher in BIT^® compared with micropropagation in liquid or semisolid media. In turn, the total content of steviol glycosides produced was also higher in bioreactors. The usefulness of BIT^® to produce plant metabolites in vitro is again demonstrated, even if …