The aim of the study was to determine the yield and chemical composition of milk from TMR-(group I) and pasture-fed Simmental cows (group II). The study was conducted with second and third lactation Simmental cows between 30 and 200 days of lactation. The present research showed that compared to TMR feeding, the use of summer pasture feeding and proper supplementation with high-energy feeds allow for higher milk yield and higher nutritive value of the milk. Compared to TMR-fed cows (group I), milk from pastured cows (group II) was characterized by a more beneficial composition of protein fractions, and a higher content of α-lactalbumin, β-lactoglobulin and lactoferrin. It also contained more vitamins A and E, calcium, magnesium and iodine, and had a significantly (P≤0.05) lower cholesterol content. The milk of cows from group II contained over twice as much CLA (1.59% of all acids) and 35% more n-3 PUFA, which resulted in a more beneficial n-6/n-3 fatty acids ratio of 2.88. In addition, this milk contained significantly (P≤0.05) less saturated fatty acids (SFA) and significantly more (P≤0.05) mono-(MUFA) and polyunsaturated fatty acids (PUFA). Consequently, the MUFA:SFA and PUFA:SFA ratios in this group were more favourable at 0.448 and 0.066, respectively. Also the content of desirable fatty acids (DFA) with hypocholesterolemic effects was higher in group II, which resulted in a more beneficial DFA:OFA ratio of 0.8 in this group. In conclusion, the use of summer pasture feeding and a proper supplemented feeding ration in Simmental cows with high-energy feeds allow for high milk yield and high nutritive value of the milk.