Recellularization of Porcine Acellular Muscle Matrix Scaffolds

The fields of tissue engineering and regenerative medicine are poised to provide innovative solutions for patients suffering from the loss of functional tissue to injury and/or disease. Current methods for repairing damaged skeletal muscle are inadequate as they require a patient’s healthy tissue to be harvested in an attempt to repair or replace damaged tissue. A preferable alternative would be to use the tissue engineering paradigm and combine a suitable biomaterial with a source of patient-specific myogenic cells to generate a construct that could facilitate the repair/regeneration of the tissue. In concurrent work, we are producing and characterizing a novel biomaterial scaffolding system we refer to as Porcine Acellular Muscle Matrix (PAMM), which is produced through the decellularization of sheets of porcine skeletal muscle. We hypothesized that PAMM scaffolds that are lyophilized, rehydrated, and sterilized are biocompatible and can support the growth and differentiation of different populations of myogenic cells. Our results demonstrate that PAMM scaffolds can be efficiently recellularized with murine C2C12 myoblasts. Histological analysis of recellularized PAMM scaffolds shows that the seeded myoblasts penetrate the entire width of the scaffold, and the alamarBlue® viability assay confirms the viability of the cells in/on the scaffold. Thus, the processing, lyophilization, rehydration, sterilization, and cell seeding procedures are adequate for us to begin using PAMM scaffolds to 1) test the myogenic potential of different stem cell populations in a three-dimensional in vitro model and 2) test the ability of unseeded or cell-seeded PAMM scaffolds to support muscle regeneration in vivo