Decellularized extracellular matrix (dECM) is designed to remove cells that cause immune rejection and retain the original tissue structure and composition. Since its structure and composition are similar to the original tissues and organs, it has attracted extensive attention in tissue engineering and biomedicine applications, and has become a promising tissue engineering material. dECM can be easily obtained from tissues and organs by appropriate decellularization methods. Here, we summarized the commonly used decellularization methods and reviewed the sterilization, cross-linking and storage methods of decellularized scaffold. In addition, we summarized the latest applications and developments of dECMs obtained from different tissues/organs in tissue engineering and biomedicine. Finally, we discussed the present challenges of dECM biomaterials and prospected future perspectives. With the development of tissue engineering and regenerative medicine technology, dECM biomaterials are expected to become a gold scaffold in the field of biomedicine and will receive wide applications.