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(1) Background: Gender-based violence has no geographical, personal, or social boundaries. It constitutes a serious public health problem that affects the entire society. This research aims to identify and compare the level of ambivalent sexism in Spanish and Colombian university students and its relationship with sociodemographic factors. Ambivalent sexism, developed by Glick and Fiske (1996), is considered a new type of sexism since, for the first time, it combines negative and positive feelings that give rise to hostile and benevolent sexism, maintaining the subordination of women through punishment and rewards. (2) Methods: The methodology consisted of the application of the validated Spanish version of the Ambivalent Sexism Inventory (ASI) to a sample of 374 students in their final academic year of the Law program, of which 21.7% were students at the University of Santander (Bucaramanga, Colombia), 45.5% at the University Rey Juan Carlos (Madrid, Spain), and the remaining 32.9% at the University of Seville (Seville, Spain). (3) Results: A high level of ambivalent sexism is reported in Colombian students nowadays. In the two countries. there are similarities (e.g., the great weight of religion and the variation in attitudes towards sexism in people who identify themselves as women, compared to male or students consulted that prefer not to answer) and differences (e.g., absence in Colombia of gender-specific legislation, low number of students who have received gender education in Spain). (4) Conclusions: These findings may contribute to the construction of laws that take into account the particular problems of women and the development of educational programs on gender that are offered in a transversal and permanent way and that take into account cultural factors and equity between men and women as an essential element in the training of future judges who have the legal responsibility to protect those who report gender violence.