The skin is the largest organ in the human body, and skin defects are very common. Skin flap transplantation is the best treatment for serious wound defects, and donor site tissues are always sacrificed during this process. Decellularized biomaterials, derived mainly from various nonautologous organs and tissues, have promising applications in tissue engineering and repair of wound defects. To date, decellularized mesothelium, intestine, amniotic membrane, dermis, and skin flaps have been developed and applied for skin coverage in animal models and clinical practice. In this review, we discuss recent advances in decellularized biomaterials for skin substitutes and future perspectives. © 2019 Wiley Periodicals, Inc. J Biomed Mater Res Part A: 107A: 1849–1859, 2019.