Hemp seed and flax seed are natural sources of various bioactive compounds, including n-3 polyunsaturated fatty acids and tocopherols. The present study aimed to evaluate the effects of hemp seed from the non-psychoactive variety Futura, non-extruded flax seed from the variety Floral, and their combination on the performance of laying hens as well as on the quality and nutritional profile of the eggs produced. A total of 240 Lohmann Brown laying hens were randomly allocated to 4 dietary treatments, with 6 replicate cages containing 10 hens each. The first control diet contained no seed supplementation. The second and third diets were supplemented with 4 % of flax seed and 4 % of hemp seed, respectively. The fourth diet included a combination of both seeds (each at 4 %). The inclusion of flax seed significantly increased both egg production and egg mass (p < 0.001). The lowest feed conversion ratio was observed in the second group, which received flax seed supplementation alone. The addition of hemp seed increased feed consumption per egg (p = 0.005) and total egg mass (p < 0.001). All the seed-supplemented groups exhibited increased egg weights. Hemp seed supplementation markedly increased yolk γ-tocopherol concentrations (p < 0.001) and reduced total cholesterol (p = 0.042) and malondialdehyde levels in fresh yolks (p = 0.003), indicating a potential improvement in oxidative stability. The combined supplementation with hemp seed and flax seed significantly increased the total polyunsaturated fatty acid content (p = 0.004), including both n-6 (p < 0.022) and n-3 (p = 0.001) polyunsaturated fatty acids. Among the n-3 polyunsaturated fatty acids, the levels of α-linolenic and eicosapentaenoic acids were elevated. These changes contributed to an improved n-6/n-3 ratio, which was significantly affected by both hemp seed and flax seed supplementation (p < 0.001). All dietary treatments significantly improved the lipid quality indices (p < 0.001). In conclusion, both hemp seed and flax seed can be considered valuable functional components of feed mixtures for laying hens, primarily due to their beneficial effects on the performance of laying hens as well as on the quality and nutritional profile of the eggs produced.