Despite its initial appearance as a straightforward bacterial infection, urinary tract infection (UTI) presents a perplexing challenge, as 20-30% cases recur or relapse despite antibiotic treatment. While the emergence of antimicrobial resistance undoubtedly contributes to some of these treatment failures, the phenomenon of recurring UTIs, even when appropriate antibiotics are administered, remains enigmatic. This disparity likely arises from intricate and multifaceted aspects of the interplay between the pathogen and its host. Simultaneously, the significance of the healthy uromicrobiome in this context remains unclear. Our current understanding of UTI host-pathogen interactions at the urothelial surface is primarily derived from forced infection experiments conducted in mice. It is worth noting, however, that mice are not natural hosts for UTIs, and their bladder environment significantly differs from that of humans. Consequently, our research team is dedicated to constructing alternative, human-cell-based laboratory models to investigate the behavior of uropathogens and the commensals that reside within the human urinary system. In this talk, I will present recent work with 3D-UHU, our most advanced static model, which we have been using to explore the biology of infection and treatment resistance using E. coli and other common uropathogenic species. These experiments highlight diverse species- and strain-specific infection strategies, illuminating why diagnosis and treatment may fail, and uncovering strategies for improving both in the future.