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In this study the CERO 3D Incubator & Bioreactor was used for the generation of tumor spheroids from human colon cancer cell line HT29-MTX-E12. The article provides information on the development of a microfluidic 3D intestine tumor spheroid model that allows for the testing of functionalized nanoparticles and their permeation over a mucus layer. The introduction indicates that oral drug delivery is preferred, and the effectiveness of orally administered drugs depends on bioavailability. Nanoparticles are an important tool in the improvement of medical treatments, and therefore, targeted delivery and effectiveness of nanoparticulate drug delivery systems can be tested using in vitro tests.