Gamified learning environments are widely used within the educational context in the subject of Physical Education. The aim of this study was to analyze the perception of the motivational climate, satisfaction, and intrinsic motivation of university students in practical classes according to gender and the type of methodology used. A total of 76 university students aged 20.58±1.69 participated in twelve two-hour practical sessions. The sessions were developed through traditional sport games, which were divided according to the type of methodology used: traditional (TRA, n=6) and gamified (GAM, n=6). Afterwards, to detect the differences in mean scores obtained for each of the methodologies used, the following tests, if needed, were performed: a non-parametric Friedman’s test with a post-hoc Wilcoxon test for pair-wise comparisons, or a two-factor repeated measures with a post-hoc Bonferroni test. Additionally, a Receiver Operating Characteristic curve analysis was performed to determine the cut-off point of the variables studied as a function of the classification of the study participants according to gender. The results showed differences in the perception of motivational climate according to gender in the traditional instructional approach. More specifically, the score for the dimension task climate was higher (M=4.21±0.58) than that recorded for the female students (M=4.57±0.45). In addition, for intrinsic motivation, the male students showed differences between both methodologies, with a higher score in the gamified program in the dimensions competence (M=3.63±0.51) effort-importance (M=3.57±0.55) and tension-pressure (M=2.92±0.85). Finally, there were significant differences between the two methodologies for satisfaction scores with the classes by gender. In each and every dimension that showed significant differences, the scores obtained by the female students were higher than those recorded for the male group in both methodologies. The application of a gamified intervention emerges as a powerful tool that could increase the motivation and satisfaction of students, favoring the teaching-learning process.