A Method for Fabricating Long Decellularized Scaffolds from Skeletal Muscle.

Skeletal muscle tissue engineering (SMTE) is a rapidly evolving field with applications spanning regenerative medicine, disease modeling, drug screening, and biohybrid robotics. Effective SMTE requires scaffolds that reproduce the anisotropic architecture and mechanical properties of native muscle while supporting macroscale tissue formation. Decellularized tissues are strong candidates; however, existing approaches face key limitations. Whole-muscle decellularization requires complex perfusion systems and often fails to fully clear the tissue core. In contrast, minced-tissue processing completely disrupts native architecture and necessitates technically demanding reconstruction. As a result, producing long, continuous scaffolds needed to model or restore physiologically relevant muscle units remains challenging. Here, we present an intermediate strategy that enables the fabrication of long, aligned scaffolds from native muscle bundles. Bundles exceeding 10 cm in length were dissected to preserve native alignment and subjected to mild detergent-based decellularization, achieving efficient removal of cellular material while maintaining extracellular matrix structure and mechanics. The resulting scaffolds supported myogenic cell adhesion, proliferation, and differentiation, demonstrating their suitability for in vitro muscle tissue culture. This accessible approach provides a straightforward route to generate macroscale, structurally faithful skeletal muscle scaffolds, bridging the gap between whole-muscle and minced-tissue decellularization methods.