Utilization of All-Chitin Composite Films for High-Density Three-Dimensional Cell Cultivation

In this study, the potential of an all-chitin composite (AChC) film, prepared by dispersing and mixing high-crystalline scaled-down chitin nanofibers (SD-ChNFs) and low-crystalline chitin nanoparticles (ChNPs) in aqueous acetic acids, was evaluated for the formation of tissue-like cellular networks using a layered culture approach. The AChC film exhibited strong adhesion properties that enabled human cervical cancer (HeLa) cells to form a cellular network structure, whereas such adhesion and network formation were not observed with the individual components, SD-ChNFs and ChNPs. Furthermore, the cellular network structure was found to bridge the gaps between films, establishing three-dimensional (3D) connectivity. These findings demonstrate that AChC films serve as highly suitable functional biomaterials for constructing 3D tissue-like structures.