Advanced Three-Dimensional Cell Culture Models for Investigating Enterohaemorrhagic Pathogenesis and Shiga Toxin-Mediated Injury.

Enterohemorrhagic (EHEC), a pathotype within the Shiga toxin-producing (STEC) group, is a major etiological agent of severe gastrointestinal illness and life-threatening sequelae, including hemolytic uremic syndrome. Although insights into EHEC pathogenesis have been gained through traditional 2D cell culture systems and animal models, these platforms are limited in their ability to recapitulate human-specific physiological responses and tissue-level interactions. Recent progress in three-dimensional (3D) cell culture systems, such as spheroids, organoids, and organ-on-a-chip (OoC) technologies, has enabled more physiologically relevant models for investigating host-pathogen dynamics. These advanced platforms offer improved modeling of human tissue architecture, cellular heterogeneity, and microenvironmental complexity, thereby providing novel perspectives on Shiga toxin-mediated damage in intestinal and renal systems. This review synthesizes current applications of 3D culture systems in EHEC research, critically evaluates their advantages and limitations, and outlines future directions for enhancing mechanistic understanding and translational relevance in STEC-associated disease modeling.