Abstract

Cultured meat has emerged as a promising approach to enhancing global food sustainability. Current production methods, however, rely mainly on single-cell-type culture, which fails to replicate the complex composition of conventional meat. In this study, a scaffold-free co-culture system was established to generate structured microtissue comprising highly differentiated muscle and fat cells. Co-cultured spheroids with the ratios of piscine adipose-derived stem cells (PADSCs) to piscine satellite cells (PSCs) being 5:5, 7:3 and 9:1 were investigated, and the result showed that all ratios remained robust and highly viable throughout the differentiation process and successfully induced the formation of bionic tissue containing both myotubes and adipocytes. Based on transcriptome data, the cluster analysis indicated that co-cultured spheroids (the ratio of PADSCs to PSCs was 7:3) were similar to the PADSC spheroids, and the expression levels of cell cycle-related and extracellular matrix-related genes were biased towards those of PADSC spheroids. These results indicated that these co-cultured spheroids held promise as building blocks for further tissue assembly and offered a promising approach for scaling up the production of structured cultured fish meat.