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 21-d periods. Dietary treatments consisted of a basal diet (Control; no fat supplement), and fat-supplemented diets containing fish oil (FO; salmon oil; 500 g/d/cow) and FO (250 g/d/cow) + hydrogenated palm oil (PO; palm oil; 250 g/d/cow). Milk collected on d 21 was pooled from 3 cows within the same treatment and period and made into cheese. Three cheeses per treatment per period at 14 d of aging were used for sensory evaluation. Cheese color measurements were determined according to the CIE L*, a*, and b* LAB color system. Sensory evaluation considered the following attributes: color homogeneity, holes, overall odor, ripe cheese odor, cow milk odor, salty, acid, bitter, overall flavor, ripe cheese flavor, sharpness, toughness, graininess, screeching, moisture and greasiness. Except for milk lactose yield, milk and cheese components were not affected by dietary treatments. Milk and cheese contents of C6:0, C8:0, C10:0 and C14:0 and atherogenicity index were lower (P < 0.05) with FO and FOPO diets than control diet. Compared with control and FOPO, FO increased (P < 0.05) C18:1 trans 11 and C22:6n3 content in milk and cheese. The color obtained from cheeses elaborated from cows given FO resulted in a more (P < 0.05) intense yellowness than control and FOPO. Compared with control and FOPO, FO resulted in higher (P < 0.05) notes for color homogeneity, whereas in comparison with control and FO, FOPO resulted in higher (P < 0.05) tough texture. 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. This study was sponsored by a research grant from FONDECYT 11121142 (Fondo Nacional de Desarrollo Científico y Tecnológico, Chile).