Seeding and Recellularization of Porcine Acellular Muscle Matrix Biomaterials with Adipose-Derived Mesenchymal Stem Cells and C2C12 Myoblasts

The ability of skeletal muscle to repair itself via regenerative mechanisms is limited to instances where tissue damage is relatively small. When volumetric muscle loss occurs, the regenerative capacity of skeletal muscle is exceeded. This results in a permanent loss of muscle volume and function. Current strategies to replace or repair such damage are inadequate. The goal of this project is to develop natural biomaterials that facilitate the engineering and/or regeneration of skeletal muscle tissue by providing a myoinductive environment for seeded and/or infiltrating cells. We hypothesized that scaffolds and hydrogels composed of porcine acellular muscle matrix (PAMM) could be efficiently recellularized and support myogenic differentiation. Here, we describe the production and characterization of PAMM scaffolds and gels. Histological analyses, DNA content measurement, and scanning electron microscopy show that porcine skeletal muscle tissue can be effectively decellularized and processed into both a sheet-like scaffold and a hydrogel. We also demonstrate that PAMM biomaterials can be recellularized with murine adipose-derived mesenchymal stem cells and the C2C12 myoblast cell line. These results demonstrate the potential for PAMM biomaterials to be employed in tissue-engineering- and regenerative-medicine-based strategies for repairing volumetric muscle loss