Plant cell cultures of Silybum marianum L. are studied under different classes of elicitors including heavy metal ions (Ag+), polysaccharides (yeast extract and chitosan), and plant response signaling compounds (salicylic acid) in order to enhance silybin production. Remarkably, all elicitors enhanced the accumulation of silybin compared to the control experiment, but the highest total silybin content was achieved in cell suspensions elicited with yeast extract at 0.5 μg.ml−1 producing a total silybin yield at 0.15 mg.g−1 cells dry weight after 2 days of elicitation for a total cell dry weight of 1.49 g. The lowest silybin accumulation belongs to chitosan treatment producing maximum silybin content and dry weight of 0.038 mg.g−1 DW and 1.19 g, respectively. These results are promising to establish a proof of concept using yeast extract as a viable elicitor option to produce silybin in tissue culture. Practical applications Optimization of elicitor‐induced cell culture production of secondary metabolites is important to develop a proof concept that can be easily scaled‐up and stand as an economically viable solution to meet the increasing demand for health and food supplements. In fact, the production of secondary metabolites from tissue cultures permits space and resource optimization, protects endangered endemic plants, and contributes to food/health security and sustainable development.