Decellularized tissue-derived extracellular matrices (ECM) and methods of generating and using same are provided. The method of generating a decellularized matrix includes the steps of: (a) subjecting the tissue to washes and a hypertonic buffer (b) subjecting the tissue to an enzymatic proteolytic digestion with an enzyme such as trypsin and (c) removing all cellular components from the tissue using a detergent solution which includes Triton-X-100 and ammonium hydroxide. Specifically, there is provided a decellularized myocardium-derived matrix which is completely devoid of all cellular components and hence non-immunogenic in a subject, exhibits suitable structural and mechanical properties for cardiac tissue engineering or replacement therapy of damaged cardiac tissue, and is capable of remodeling upon seeding of cells.
