This assessor-blinded, randomized controlled superiority trial investigated the efficacy of the 10-week Nordic Hamstring exercise (NHE) protocol on sprint performance in football players. Thirty-five amateur male players (age: 17–26 years) were randomized to a do-as-usual control group (CG; n = 17) or to 10-weeks of supervised strength training using the NHE in-season (IG; n = 18). A repeated-sprint test, consisting of 4 × 6 10 m sprints, with 15 s recovery period between sprints and 180 s between sets, was conducted to evaluate total sprint time as the primary outcome. Secondary outcomes were best 10 m sprint time (10mST) and sprint time during the last sprint (L10mST). Additionally, peak eccentric hamstring strength (ECC-P_HS) and eccentric hamstring strength capacity (ECC-CAP_HS) were measured during the NHE. Ten players were lost to follow-up, thus 25 players were analyzed (CG n = 14; IG n = 11). Between-group differences in mean changes were observed in favor of the IG for sprint performance outcomes; TST (−0.649 s, p = 0.056, d = 0.38), 10mST (−0.047 s, p = 0.005, d = 0.64) and L10mST (−0.052 s, p = 0.094, d = 0.59), and for strength outcomes; ECC-P_HS (62.3 N, p = 0.006, d = 0.92), and ECC-CAP_HS (951 N, p = 0.005, d = 0.95). In conclusion, the NHE showed small-to-medium improvements in sprint performance and large increases in peak eccentric hamstring strength and capacity. Trial Registration Number: NCT02674919