High density culture of bovine embryonic stem cell derived mesenchymal cells on edible scaffolds for structured cultivated meat

Abstract

Developing structured cultivated meat requires integrated solutions that combine scalable cell sources with edible, foodgrade materials capable of supporting highdensity growth and differentiation. Here, we evaluate bovine mesenchymal stem cells derived from embryonic stem cells (ESCderived iMSCs) as a scalable adipogenic cell source and develop an integrated workflow combining these cells with edible plantbased scaffolds for structured biomass generation. Cell identity and functionality were assessed using transcriptomic, morphological, gene expression, flow cytometric, and adipogenic differentiation analyses, in both adherent and suspension culture systems. In parallel, lentil, pea, and soy-based scaffold formulations were screened for cell attachment, proliferation, and biomass accumulation. Soybased scaffolds supported uniform cell distribution and robust growth and outperformed lentil-based scaffolds. Under dynamic culture conditions, bovine iMSCs cultured on soy-based scaffolds achieved highdensity growth, showing biomass accumulation (cell wet weight/scaffold wet weight) reached an average cell wet weight to scaffold wet weight ratio of 15% within three days. Cultures demonstrated active glucose metabolism and retained adipogenic differentiation capacity, confirmed by lipid accumulation and positive oil red O staining. These findings demonstrate an integrated cell-scaffold platform for rapid threedimensional biomass generation. This approach supports the development of a cell culture strategy for structured cultivated meat by combining defined cell sources with foodgrade scaffold technologies to improve scalability, structure, and nutritional relevance. HighlightsO_LIBovine ESC-derived iMSCs enable scalable adipogenic cell production C_LIO_LIEdible soy-based scaffolds support 3D attachment and biomass accumulation C_LIO_LIDynamic culture achieved [~]15% cell wet weight fraction within 3 days C_LIO_LIiMSCs retained adipogenic differentiation capacity on edible scaffolds C_LIO_LIIntegrated cell-scaffold culture supports structured cultivated meat prototypes C_LI