Maximum running speed and acceleration are essential components in many sports. The identification of specific training protocols to maximize sprint speed would be useful knowledge for coaches and players. The purpose of this study was to determine the effect of a high-speed treadmill (HST) with the use of a body weight support (BWS) system in a 6-week sport acceleration program (SAP) on female soccer athlete's 40-yard sprint time and maximal isometric knee flexor and extensor strength. Two treatment groups and one control group were created. Both treatment groups participated in a 12-session SAP. The first treatment group (n = 12) used a BWS system while running on a HST; the second group (n = 12) used a standard treadmill (ST) with no BWS system. The participants of the control group (n = 8), NT, did not participate in a sports acceleration program and did not alter their exercise routines outside of the study. An analysis of covariance was performed using baseline measures as the covariate. The 40-yard sprint times for both treatment groups were shown to improve significantly compared with the control group (p < 0.001). Isometric knee flexor strength showed a greater increase in the ST group (p = 0.026) than in the other 2 groups, whereas knee extensor strengths did not show significant differences between treatment groups and control group (p > 0.05). Participants in the ST group had a much higher rate (66%) of shin splints and foot pain throughout the study than those in the HST (8%) and NT (0%) groups. These results can help high school coaches and athletes determine the optimal treadmill training regime.