Heterogeneity Governs 3D‐Cultures of Clinically Relevant Microbial Communities

Abstract The intrinsic heterogeneity of bacterial niches should be retained in vitro cultures to represent the complex microbial ecology. As a case study, mucin‐containing hydrogels ‐CF‐Mu 3 Gel ‐ are generated by diffusion‐induced gelation, bioinspired on cystic fibrosis (CF) mucus, and niche challenging current therapeutic strategies. At breathing frequency, CF‐Mu exhibits G ′ ″ equal 24 3.2 Pa, respectively. Notably, structural gradients with gradual reduction oxygen tension across its thickness (280–194 µmol L −1 ). Over culture period, steep decline concentration occurs just few millimeters below air–mucus interface Gel, similar those CF airway mucus. Importantly, distinctive features significantly influence organization antimicrobial tolerance mono‐ co‐cultures Staphylococcus aureus Pseudomonas aeruginosa that standard unable emulate. susceptibility determined corroborates mismatch efficacy treatment between planktonically cultured bacteria patients. With this example‐based research, new light is shed understanding how substrate influences behavior, paving way for improved fundamental microbiology studies more effective drug testing development.