OBJECTIVES: Urban-rural comparisons between those who maintain traditional lifestyles such as pastoralist Maasai children, and those who live in cities such as Nairobi, provide implications on how urbanization is associated with children's physical activity (PA) and sedentary behavior (SB) patterns. This study compares PA and SB volumes and patterns across different segments of the week among children in Maasai village and Nairobi city in Kenya. METHOD: A total of 261 children (11.4 ± 1.3 years) from Maasai (n = 118) and Nairobi (n = 143) participated in this cross-sectional study. Moderate- to vigorous-intensity PA (MVPA) and SB on weekdays (before, during, and after school) and weekends (morning, afternoon, and evening) were calculated using accelerometers (ActiGraph). Screen time and sleep duration were assessed using questionnaires. RESULTS: Maasai children were more physically active than Nairobi children with MVPA (min/day) of 166.6 and 81.4 for Maasai and Nairobi boys and 116.4 and 77.4 for Maasai and Nairobi girls, respectively. Our week segments analyses suggested that Maasai children were more active both in and out of school than Nairobi children. Additionally, Nairobi children spent more time watching television and playing computer games than Maasai children. There was no significant difference in sleep duration between Maasai and Nairobi children. CONCLUSION: Our findings suggest that urbanization is negatively associated with activity patterns both in and out of school in Kenyan children. This is concerning given that Kenya is currently undergoing rapid urbanization, which may lead to further reductions in PA among Kenyan children.