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Bryophytes, due to their poikilohydric nature and peculiar traits, are useful and versatile organisms for studies on metal accumulation and detoxification in plants. Among bryophytes, the liverwort Marchantia polymorpha is an excellent candidate as a model organism, having a key role in plant evolutionary history. In particular, M. polymorpha axenic cultivation of gametophytes offers several advantages, such as fast growth, easy propagation and high efficiency of crossing. Thus, the main purpose of this work was to promote and validate experimental procedures useful in the establishment of a standardized set-up of M. polymorpha gametophytes, as well as to study cadmium detoxification processes in terms of thiol-peptide production, detection and characterisation by HPLC-mass spectrometry. The results show how variations in the composition of the Murashige and Skoog medium impact the growth rate or development of this liverwort, and what levels of glutathione and phytochelatins are produced by gametophytes to counteract cadmium stress.