Human amniotic membrane (HAM) has been used as a very promising biological-based product in health centers, especially for skin and cornea wound healing applications. The excellent properties of this membrane make it a potential candidate in treatment of various skin injuries such as bedsores, burn wounds and diabetic ulcers. Such properties are cytobiocompatibility, a structure very similar to normal skin composed of extracellular matrix (ECM) proteins, various growth factors involved in normal wound healing process and antibacterial agents. HAM contains epithelial cells, fibroblasts and mesenchymal stem cells. Therefore, the successful decellularization of HAM with minimal negative effects on its ECM components is very important to avoid graft rejection and shows improved performance. To date, several approaches have been conducted for decellularization of HAM, which is mainly based on enzyme-, detergent- or mechanical procedures with various ranges of success. Here, we describe a systematic detergent based decellularization protocol as main protocol. We also explain the enzyme- and mechanical based methods as the alternative protocols for decellularization of HAM.