Engineered marbled meat through EPA-mediated co-differentiation of porcine muscle and fat progenitors without conventional hormonal induction

Introduction Intramuscular fat, known as “snowflake” pattern in pork, critically determines meat tenderness, juiciness, flavor, and overall consumer preference. Cultured meat has emerged as a promising solution to address future protein demands sustainably. However, producing cultured meat with authentic marbling remains challenging, as myocytes and adipocytes require distinct microenvironments for differentiation. Objective This study aimed to develop a co-differentiation strategy independent of conventional hormonal adipogenic cocktails (IBMX and dexamethasone) to simultaneously induce myogenesis and adipogenesis in a single culture system, thereby enabling the production of structured cultured meat with microscale muscle-fat patterning. Methods An edible κ-carrageenan-konjac glucomannan (CK) hydrogel was developed as a versatile platform for cultured meat. A 3D co-culture system was established by encapsulating porcine muscle satellite cells (pMuSCs) and fibro-adipogenic progenitors (pFAPs) within the CK scaffold, followed by differentiation induction using an eicosapentaenoic acid (EPA, C20:5n-3)-based strategy. The physicochemical properties of the resulting cultured meat induced with EPA (CM-EPA) were rigorously characterized using texture profile analysis and nutritional composition analysis, with comparisons made against horse serum-induced controls (CM-HS) and commercial streaky pork. Results The CK hydrogel exhibited tunable, biomimetic mechanical properties and a porous microstructure that supported the proliferation and differentiation of both cell lineages. Notably, the EPA-based induction strategy significantly enhanced lipid accumulation while simultaneously supporting myotube formation from pMuSCs, effectively mitigating the myogenesis inhibition associated with conventional hormonal adipogenic cocktails. The co-cultured meat exhibited significantly improved textural properties, such as hardness and chewiness, which closely resembled those of real streaky pork. Its nutritional profile also showed elevated protein and fat content relative to the CM-HS control. Conclusion This work establishes a versatile scaffold-based platform for producing structured cultured meat with integrated muscle and fat components, offering a promising strategy to replicate the textural and nutritional qualities of traditional meat.