Abstrakt
Stroma of mammalian organs including skeletal muscle is so complex in its macroscopic structure, microarchitecture and molecular composition that its detailed artificial reconstruction is out of reach. Nevertheless a removal of cells from the organ can be used to yield the extracellular matrix (ECM) mimicking many features of the original tissue complexity.
We decellularized the skeletal muscle using combination of the osmotic shock, chemical and DNase treatment and obtained the bioscaffolds free of cellular and cytoplasmic components. Moreover, the scaffold ECM reflected light microscopic organization of the muscle coverings, blood vessels and peripheral nerves and also retained 3D organization of basal laminas and collagen fibrils as confirmed by transmission electron microscopy.
Laminin and collagen type IV remained organized within basal laminas supporting cell attachment and navigation. Recellularization of the scaffolds with C2C12 cells in vitro confirmed cytocompatibility of the decellularized ECM as well as its ability to support basic physiological cellular functions as the myoblasts were aligned with ECM microstructure.
After 12 days in vitro the samples did not show any signs of abnormalities, regression or cell death. These data strongly indicated that skeletal muscle decellularization could be considered as a promising approach in construction of bioscaffolds suitable for muscle tissue replacement.