The intestinal mucus layer remains a challenging biological barrier to cross for nanoformulations. We report the assembly of zwitterionic micelles with 2-methacryloyloxyethyl phosphorylcholine as the zwitterionic entity in the corona-forming part of the amphiphilic block copolymers. These ca. 200 nm micelles do not have inherent cytotoxicity in Caco-2 cells for a 24 h incubation time. The time-resolved crossing of the mucus layer in Caco-2/HT29-MTX-E12 co-cultures of these spherical micelles with narrow polydispersity is shown.