Sprouts from germinated seeds of edible herbaceous plants are richer in bioactive compounds and micronutrients than mature plants. In recent years, the consumption of sprouted seeds in diets and the interest in exploring vegetable sprout's potential beneficial effects on human health has increased. In this work, the five-day-old sprouts of Cannabis sativa L. cultivar “Futura 75” were investigated and characterized. Hemp sprouts were obtained by germination tests under standard and enrichment conditions using the microalgal strain Chlorella sp. C2 and the bacterium Sphingomonas sp. Can_S11. The biochemical properties of hemp sprouts were analyzed for total polyphenol and flavonoid content and antioxidant activities (DPPH, ABTS, FRAP, ORAC, and Fe2+-chelating activity) by in vitro assays. A significant increase in phenolic compounds and greater antioxidant and anti-radical activities were observed for hemp sprouts obtained under enrichment conditions (dry microalgal biomass C2 and the bacterium Can_S11). Additionally, these sprouts and C. sativa seeds demonstrated enhanced protective effects on inflamed A549 cells, effectively mitigating the TNF-α induced changes and reducing the expression of IL-8 and COX-2. Overall, PGP (Plant Growth Promoting) microorganisms and microalgae represent a good strategy to promote the development of hemp sprouts and increase their phytochemical content, and antioxidant and anti-inflammatory activity.