A Chitosan-Based Micromesh Platform for Enhanced Myoblast Alignment and Three-Dimensional Tissue Formation in Cultured Meat

As global demand for meat continues to rise, cultured meat technology is emerging as a sustainable solution to solve future meat shortages. While various three-dimensional cell culture methods have been developed, those without scaffolds produce tissues that are difficult to handle and lack the layered structure of natural tissues, making them unsuitable for cultured meat production. Conversely, scaffold-based methods are easier to manage but result in significant scaffold material inclusion in the final product. Additionally, creating tissue-like layered structures has been labor-intensive.<br/><br/>In this study, we developed a cultured meat platform utilizing edible micromesh sheets composed of chitosan. These micromesh sheets form a high-porosity, diamond-shaped micromesh sheet. Our cultivation method involves first adhering cells to the microscale mesh sheet, followed by the formation of a cell sheet as proliferating cells fill the mesh openings. In this approach, cells within the mesh openings primarily rely on cell-cell adhesion for survival, mimicking the environment of cell growth in vivo and differing from cells attached to conventional plastic substrates. Cells dependent on intercellular adhesion can easily form three-dimensional structures, enabling the facile production of thick cell sheets through this mesh culture method.<br/><br/>Using this carrier, we successfully cultivated thick cell sheets of C2C12 cell line, a mouse myoblast cell line. We demonstrated that the cells align uniaxially along the long axis of the diamond-shaped openings, exhibiting characteristic alignment properties. Simulating this alignment pattern is crucial for achieving physiologically relevant cell sheets for potential applications in cultured meat production or regenerative medicine.