Abstract

Cultured meat is an emerging technology to produce edible muscles using animal cells, scaffolds, and bioreactors, avoiding the slaughter of farmed animals. Since muscle cells tend to grow disorganized, the use of microstructuring technologies to produce scaffolds is essential to align functional muscle structures. In this work, an edible scaffold (film) with parallel microchannels formulated with marine biopolymers was made. The effect of parallel microchannels on the behavior of muscle cells and gene expression involved in fusion and myogenesis were assessed. Our results show that the array of microchannels allow the formation of parallel, elongated, and multinucleated structures, like a muscle bundle. In addition, the glycolytic metabolism increases when cells grew on the microchannels. Significant correlations between gene expressions of Myomaker and MyoD, between Myomixer and Myosin heavy chain, and between Myogenin and Myosin heavy chain were found and associated with the microstructure effect. An inferred model of myogenic progression was proposed and validated identifying proteins by immunofluorescence on cells seeded on the microstructured film-scaffolds. This model allows to understand the role of myogenic markers in the production of cultured meat using microstructured scaffolds.