Antimicrobial resistance (AMR) is a growing threat that causes over 700,000 deaths per year worldwide. The goal of the current multicenter, cross-sectional study was to identify the knowledge and practice gaps in antimicrobial stewardship among Egypt’s undergraduate medical students. Nine-hundred and sixty-three participants (375 male) from 25 medical schools responded to our self-administered questionnaire. Overall, the majority of students (96%) exhibited fair/satisfactory knowledge and attitude scores towards AMR. However, the most common misconceptions were that skipping one or two antimicrobial doses does not contribute to AMR (43%) and that antimicrobials are the drug of choice for the treatment of sore throat (38.8%). About 36% of the students thought that bacteria cause common cold and influenza. In terms of practices, about 62% of the students reported taking antibiotics when they have cough or sore throat and saving the remaining antibiotic for the next time. About 48% of the students reported that when they start feeling better, they stop the antibiotic course, and 41% stated that they often/sometimes discard the remaining leftover or share the leftover antibiotics with their friends. Interestingly, males had more frequently poorer levels of knowledge than females (p = 0.02). Moreover, students in the clinical science years (p < 0.001), living in urban areas (p= 0.02) or Cairo (p < 0.01) reported better practices than their counterparts. Educational programs about antimicrobial stewardship and the role of healthcare professionals in preventing AMR should be introduced early in medical curricula. Further, active educational techniques as clinical scenarios that simulate clinical settings and interactive learning workshops would be more efficient teaching methods.