This study compared the impact of two polarized training programs (POL) on aerobic capacity in well-trained (based on maximal oxygen uptake and training experience) female cyclists. Each 8-week POL program consisted of sprint interval training (SIT) consisting of 8–12 repetitions, each lasting 30 seconds at maximal intensity, high-intensity interval training (HIIT) consisting of 4–6 repetitions, each lasting 4 minutes at an intensity of 90–100% maximal aerobic power, and low-intensity endurance training (LIT) lasting 150–180 minutes with intensity at the first ventilatory threshold. Training sessions were organized into 4-day microcycles (1st day—SIT, 2nd day—HIIT, 3rd day—LIT, and 4th day—active rest), that were repeated throughout the experiment. In the first POL program, exercise repetitions during SIT and HIIT training were performed with freely chosen cadence above 80 RPM (POLFC group, n = 12), while in the second POL program with low cadence 50–70 RPM (POLLC group, n = 12). Immediately before and after the 8-week POL intervention, participants performed an incremental test to measure maximal aerobic power (Pmax), power achieved at the second ventilatory threshold (VT2), maximal oxygen uptake (VO2max), maximal pulmonary ventilation (VEmax), and gross efficiency (GE). Moreover, participants performed VO2max verification test. Analysis of variance showed a repeated measures effect for Pmax (F = 21.62; η2 = 0.5; p = 0.00), VO2max (F = 39.39; η2 = 0.64; p = 0.00) and VEmax (F = 5.99; η2 = 0.21; p = 0.02). A repeated measures x group mixed effect was demonstrated for Pmax (F = 4.99; η2 = 0.18; p = 0.03) and VO2max (F = 6.67; η2 = 0.23; p = 0.02). Post-hoc Scheffe analysis showed that increase in Pmax were statistically significant only in POLLC group. The Friedman test showed that VT2 differed between repeated measures only in the POLLC group (χ2 = 11; p = 0.001; W = 0.917). In conclusion, it was found that POL program where SIT and HIIT were performed at low cadence was more effective in improving aerobic capacity in well-trained female cyclists, than POL with SIT and HIIT performed at freely chosen cadence. This finding is a practical application for athletes and coaches in cycling, to consider not only the intensity and duration but also the cadence used during various interval training sessions.