The configuration of a biologically fertile substrate for edible plant growth during long-term manned missions to Mars constitutes one of the main challenges in space research. Mars regolith amendment with compost derived from crew and crop waste in bioregenerative life support systems (BLSS) may generate a substrate able to extend crew autonomy and long-term survival in space. In this context, the aim of our work was threefold: first, to study the geochemistry and mineralogy of Mojave Mars Simulant (MMS-1) and the physico-chemical and hydraulic properties of mixtures obtained by mixing MMS-1 and green compost at varying rates (0:100, 30:70, 70:30, 100:0; v:v); secondly, to evaluate the potential use of MMS-1 as a growing medium of two lettuce (Lactuca sativa L.) cultivars; thirdly, to assess how compost addition may impact on sustainability of space agriculture by exploiting in situ resources. MMS-1 is a coarse-textured alkaline substrate consisting mostly of plagioclase, amorphous material and secondarily of zeolite, hematite and smectites. Although it can be a source of nutrients, it lacks organic matter, nitrogen, phosphorus and sulphur, which may be supplied by compost. Both cultivars grew well on all mixtures for 19 days under fertigation. Red Salanova lettuce produced a statistically higher dry biomass, leaf number and area than Green Salanova. Leaf area and plant dry biomass were the highest on 30:70 simulant:compost mixture. Nevertheless, the 70:30 mixture was the best substrate in terms of pore-size distribution for water-plant relationship and the best compromise for plant growth and sustainable use of compost, a limited resource in BLSS. Many remaining issues warrant further investigation concerning the dynamics of compost production, standardisation of supply during space missions and representativeness of simulants to real Mars regolith.