Saffron (Crocus sativus L.) is cultivated worldwide. Its stigmas represent the highest-priced spice and contain bioactive compounds beneficial for human health. Saffron cultivation commonly occurs in open field, and spice yield can vary greatly, from 0.15 to 1.5 g m−2, based on several agronomic and climatic factors. In this study, we evaluated saffron cultivation in soilless systems, where plants can benefit from a wealth of nutrients without competition with pathogens or stresses related to nutrient-soil interaction. In addition, as plant nutrient and water uptake can be enhanced by the symbiosis with arbuscular mycorrhizal fungi (AMF), we also tested two inocula: a single species (Rhizophagus intraradices) or a mixture of R. intraradices and Funneliformis mosseae. After one cultivation cycle, we evaluated the spice yield, quality (ISO category), antioxidant activity, and bioactive compound contents of saffron produced in soilless systems and the effect of the applied AMF inocula. Spice yield in soilless systems (0.55 g m−2) was on average with that produced in open field, while presented a superior content of several health-promoting compounds, such as polyphenols, anthocyanins, vitamin C, and elevated antioxidant activity. The AMF symbiosis with saffron roots was verified by light and transmission electron microscopy. Inoculated corms showed larger replacement corms (+50% ca.). Corms inoculated with R. intraradices performed better than those inoculated with the mix in terms of spice quality (+90% ca.) and antioxidant activity (+88% ca.). Conversely, the mixture of R. intraradices and F. mosseae increased the polyphenol content (+343% ca.). Thus, soilless systems appeared as an effective alternative cultivation strategy for the production of high quality saffron. Further benefits can be obtained by the application of targeted AMF-based biostimulants.