Both strength and endurance training have several positive effects on aging muscle and physical performance of middle-aged and older adults, but their combination may compromise optimal adaptation. This study examined the possible interference of combined strength and endurance training on neuromuscular performance and skeletal muscle hypertrophy in previously untrained 40–67-year-old men. Maximal strength and muscle activation in the upper and lower extremities, maximal concentric power, aerobic capacity and muscle fiber size and distribution in the vastus lateralis muscle were measured before and after a 21-week training period. Ninety-six men [mean age 56 (SD 7) years] completed high-intensity strength training (S) twice a week, endurance training (E) twice a week, combined training (SE) four times per week or served as controls (C). SE and S led to similar gains in one repetition maximum strength of the lower extremities [22 (9)% and 21 (8)%, P<0.001], whereas E and C showed minor changes. Cross-sectional area of type II muscle fibers only increased in S [26 (22)%, P=0.002], while SE showed an inconsistent, non-significant change [8 (35)%, P=0.73]. Combined training may interfere with muscle hypertrophy in aging men, despite similar gains in maximal strength between the strength and the combined training groups.