The objective of the present study was to evaluate the effect of dietary supplementation of fish oil (FO) alone or in combination with hydrogenated palm oil (FOPO) on the fatty acid (FA) profile of milk and cheese from dairy cows, and the sensory characteristics of cheese. Nine Holstein cows (173 ± 21 DIM) were used in a replicated 3. × 3 Latin square design with three 21 d periods. Except for milk lactose yield, milk and cheese components were not affected by dietary treatment. Cheese elaborated from milk when cows were supplemented with FO had a more intense yellowness and higher notes for colour homogeneity than cheese elaborated from milk when cows were on control and FOPO treatments. FOPO treatment resulted in cheese with higher tough texture than control and FO treatments. Milk and cheese contents of C6:0, C8:0, C10:0 and C14:0 and atherogenicity index were lower with FO and FOPO treatments than with control. Compared with control and FOPO, FO treatment increased VA and DHA content in milk and cheese. In conclusion, supplementation of dairy cow diets with FO alone or in combination with hydrogenated palm oil can enhance the FA profile of milk and cheese without deleterious effects on sensory characteristics of cheese.