Haach, V.; Silveira, K. R. D.; Peixoto, M. A.; Sa, A. P. P.; Gressler, V.; Feddern, V.; Ibelli, A. M. G.; Silva, L. P.; Bastos, A. P.

Cultured meat aims to replicate the sensory and functional properties of conventional meat by developing structured muscle tissue using cell culture. This study focuses on the culture of chicken embryonic and mesenchymal stem cells (MSCs) to derive muscle, and fat, optimizing conditions for differentiation and integration. We utilized monolayer and three-dimensional microcarrier-based cultures to produce muscle fibers and adipocytes while maintaining the extracellular matrix (ECM) integrity essential for tissue cohesion. Key pluripotency and myogenic markers (e.g., cOCT4, cMYOD, cMYH1E) were analyzed during differentiation, revealing dynamic gene expression patterns that underscore myogenesis. Myoblast differentiation into mature myotubes demonstrated decreased cPAX7 and increased cMYMK, confirming lineage commitment and muscle fiber formation. Adipogenesis was induced in MSCs using food-grade lecithin, which activated PPAR{gamma}, C/EBP, and FABP4, resulting in robust lipid droplet accumulation. To scale production, microcarriers facilitated cell proliferation, while transglutaminase-based stabilization enabled the formation of three-dimensional tissue structures comparable to native meat. Our findings highlight advances in culture protocols, genotypic and phenotypic expression analyses of multinucleated chicken muscle and adipocyte cells for cultured meat production.