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Biomaterial choice is an essential step during the development tissue engineering and regenerative medicine (TERM) applications. The selected biomaterial must present properties allowing the physiological-like recapitulation of several processes that lead to the reestablishment of homeostatic tissue or organ function. Biomaterials derived from the extracellular matrix (ECM) present many such properties and their use in the field has been steadily increasing. Considering this growing importance, it becomes imperative to provide a comprehensive overview of ECM biomaterials, encompassing their sourcing, processing, and integration into TERM applications. This review compiles the main strategies used to isolate and process ECM-derived biomaterials as well as different techniques used for its characterization, namely biochemical and chemical, physical, morphological, and biological. Lastly, some of their applications in the TERM field are explored and discussed. [Display omitted] •The main methods for ECM isolation from tissue, organs, and cells and subsequent decellularization are reviewed.•Techniques for its biochemical, physical, morphological and biological characterization are described.•Examples of the application of ECM-derived biomaterials are briefly compiled and discussed.•Future focal points of research related with ECM-derived biomaterials are anticipated.