Chitosan-sodium alginate-collagen/gelatin three-dimensional edible scaffolds for building a structured model for cell cultured meat

Cell cultured meat (CCM) production is an innovative technology that does not depend on livestock farming practices to produce meat. The construction of structured CCM requires a three-dimensional (3D) scaffold to mimic the extracellular matrix to provide mechanical support for the cells. Furthermore, the 3D scaffolds should be edible and have good biocompatibility and tissue-like texture. Here, we demonstrated a 3D edible chitosan-sodium alginate-collagen/gelatin (CS-SA-Col/Gel) scaffold that can support the adhesion and proliferation of porcine skeletal muscle satellite cells, culminating in the construction of a structured CCM model. The 3D edible scaffolds were prepared by freeze-drying using electrostatic interactions between chitosan and sodium alginate. Initially, the physicochemical properties and structural characteristics of different scaffolds were explored, and the biocompatibility of the scaffolds was evaluated using the C2C12 cell model. The results showed that the 2-CSSA-Col1-Gel scaffold provided stable mechanical support and abundant adhesion sites for the cells. Subsequently, we inoculated porcine skeletal muscle satellite cells on the 2-CS-SA-Col1-Gel scaffold and induced differentiation for a total of 14 days. Immunofluorescence staining results showed cytoskeleton formation, and Western blotting (WB) and qPCR results showed upregulation of skeletal proteins and myogenic genes. Ultimately, the structured CCM model has similar textural properties (chewiness, springiness and resilience) and appearance to those of fresh pork. In conclusion, the method of constructing 3D edible scaffolds to prepare structured CCM models exhibits the potential to produce cell cultured meat.