Given the increasing development of morbidity and mortality in Clostridium difficile infection (CDI), global antibiotic resistance problems and limited alternatives to CDI prevention, it is important to identify the causes of the CDI and find new avenues to fight this infection. Unfortunately, the most common platform used in CDI research are animal models, while available in vitro models fail to ensure the viability of gut microbiota. Therefore, new in vitro model is needed for CDI testing in the gut-on-chip (GoC) model in the presence of full microbiota, as this, compared to classic animal models, provides better process control, human cell use, improved microbiota environmental simulation, reduced animal use and the ability to maintain complex and stable microbial colonies, making them a valuable tool for understanding CDI manifestation causes and developing new treatments. The aim of this project is to create a GoC microfluidic model in coculture with full gut microbiota to test the development of CDI by performing the following tasks: Create a GOC model using mucus-producing cells to model intestinal epithelium, isolate and characterize microbiota samples, and evaluate CDI effects in the GOC and microbiota model before and after antibiotic treatment is initiated. The expected results include an understanding of the role of gut microbiota composition in CDI manifestation and a functional in vitro model for further CDI research and development of therapeutic solutions.