The aim of the present study was to examine the effect of in-season strength maintenance training frequency on strength, jump height, and 40-m sprint performance in professional soccer players. The players performed the same strength training program twice a week during a 10-week preparatory period. In-season, one group of players performed 1 strength maintenance training session per week (group 2 + 1; n = 7), whereas the other group performed 1 session every second week (group 2 + 0.5; n = 7). Only the strength training frequency during the in-season differed between the groups, whereas the exercise, sets and number of repetition maximum as well as soccer sessions were similar in the 2 groups. The preseason strength training resulted in an increased strength, sprint, and jump height (p < 0.05). During the first 12 weeks of the in-season, the initial gain in strength and 40-m sprint performance was maintained in group 2 + 1, whereas both strength and sprint performance were reduced in group 2 + 0.5 (p < 0.05). There was no statistical significant change in jump height in any of the 2 groups during the first 12 weeks of the in-season. In conclusion, performing 1 weekly strength maintenance session during the first 12 weeks of the in-season allowed professional soccer players to maintain the improved strength, sprint, and jump performance achieved during a preceding 10-week preparatory period. On the other hand, performing only 1 strength maintenance session every second week during the in-season resulted in reduced leg strength and 40-m sprint performance. The practical recommendation from the present study is that during a 12-week period, 1 strength maintenance session per week may be sufficient to maintain initial gain in strength and sprint performance achieved during a preceding preparatory period.