The extracellular matrix of beef: A characterization towards a 3D scaffold for cultured meat

Scaffold development for muscle cell growth in cultured meat production requires understanding the mechanical properties of the extracellular matrix (ECM) in bovine muscle tissue. However, previous studies have focused on non-bovine samples and have altered the native ECM structure by processing it into hydrogels. This study characterizes the native ECM of bovine muscle tissues, providing reference data for designing ECM alternatives while preserving its structural and mechanical features. Protocols were optimized for thin samples (∼ 1.5 mm), corresponding to the cultured meat products currently under development. ECM was isolated from fresh bovine sirloin and tenderloin using 0.5 % SDS for decellularization. An extensive sample quality validation was performed, including cryo-SEM to visualize internal structures, Picogreen assays to quantify residual dsDNA and confirm effective cell removal, and proteomic and glycosaminoglycan analyses to verify retention of essential ECM components. Subsequent mechanical analyses included amplitude sweeps (G’, G’’, linear visco-elastic region, cross-over point) and texture analyses (total extension, maximal load). Results confirmed efficient decellularization, preserving the ECM’s structural and compositional integrity. The storage modulus (G’) was 15.2 kPa for sirloin and 12.3 kPa for tenderloin, decreasing significantly to 0.3 kPa (p < 0.01) and 1.0 kPa (p < 0.001) for their decellularized counterparts. Texture analyses revealed no significant differences, suggesting ECM primarily determines these properties. Maximal load ranged from 1.4 N to 3.1 N, while total extension varied between 9.2 mm and 11.6 mm. These findings provide reference data for scaffolds replicating natural ECM properties and establish target mechanical properties for cultured muscle fiber constructs.