Urbanization leads to significant changes in land-use types and affects various ecosystem services. While previous studies have analyzed the interaction between urbanization and ecosystem services at the national, provincial, and city scales, there is insufficient information on the spatial interaction between urbanization and ecosystem services in urban agglomerations for different development levels. To address these research gaps, this study used hot spot analysis and spatial autocorrelation models to measure the spatial interaction between urbanization and ecosystem services in China’s ten major urban agglomeration areas for the period 1995–2018. The results found that: (1) China’s urban agglomerations have experienced rapid urbanization, exhibiting a rising growth trend at all development levels of urbanization. Those grouped in the growing category exhibited the fastest growth, while those in the mature category posted the slowest growth. The spatial distributions of the urbanization indicators differ significantly. The population density showed a more pronounced clustering feature, while the proportion of urban land use was distributed mainly around the core cities. (2) Different ecosystem services exhibited varying change trends. Food supply increased significantly. Carbon storage decreased in all but two urban agglomerations (Beijing-Tianjin-Hebei and Sichuan-Chongqing). Habitat quality services also decreased in all agglomerations, except for the northern slope of the Tianshan Mountains. In terms of ecosystem service hot spots, there is distinct spatial heterogeneity. Food supply hot spots are concentrated in the plain areas, while habitat quality and carbon storage hot spots are mainly clustered in mountain and water regions. (3) Carbon storage and habitat quality are negatively correlated with the three indicators of urbanization, while food supply is positively correlated with population density and GDP density and negatively correlated with the proportion of urban land use. Urban land expansion is directly linked to the decline in ecosystem services, while population and economic growth have indirect effects. This study can contribute to understanding the correlation between urbanization and ecosystem services, and providing references for high-quality spatial development.