Scaffolds are used in the field of tissue engineering for the reconstruction of organs and tissues. These can be synthetic, made from materials such as polymers, metals, ceramics, or graphene, or natural, using macromolecules like collagen or gelatin. While useful, synthetic scaffolds often face biocompatibility issues, whereas traditional natural scaffolds can elicit insufficient cellular responses. To overcome these limitations, recent research has focused on the development of scaffolds derived from decellularized extracellular matrices (dECM). These natural-origin scaffolds are produced through a process known as decellularization, which aims to remove all cells from an organ or tissue while preserving the extracellular matrix components. Decellularization can be achieved through physical, chemical, and/or enzymatic methods, resulting in biocompatible, hypoimmunogenic scaffolds that replicate the functionality, structure, and biochemical properties of native tissue. These characteristics have led to the use of dECM in reproduction to treat infertility and enhance assisted reproductive technologies (ARTs). However, the decellularization technique is still in its early stages of development, with ongoing studies aimed at optimizing decellularization methods and clinical applications of derived materials. This chapter summarizes the basic principles of this technique and its main applications in reproduction, emphasizing the advances made to date and the challenges that remain.