Tissue engineered skin constructed by cultured seed cells provides a new way to accelerate skin wound healing and improve the quality of tissue repair. Conventional models with dermal scaffold material based on collagen hydrogels, however, have poor stability and unsatisfactory longevity. Here, we describe an improved acellular amniotic membrane as a scafford with epidermal stem cells for tissue engineering skins and observe their clinical effect on deep burn wounds. The human epidermal stem cells was isolated from the skin samples by trypsin digesting method and purified by collagen adhering method, and then seeded on the acellular amniotic membrane to form the engineered skin. The biological attachment and growth of cultured epidermal stem cells were observed. Twelve cases of deep burn wounds were applied with the engineered skin. The results showed that the epidermal stem cells adhered to the surface of acellular amniotic membrane quickly after being seeded and exhibited a high colony formation efficiency. The wounds treated with engineered skin healed rapidly with good clinical "take". The average time of wound closure in engineered skin group was shortened than that of control group. There were no obvious evidence of immunological rejection and inflammatory reaction during the observation. This indicated that the acellular amniotic membrane with good histocompatibility may be a suitable scaffold with epidermal stem cells in the construction of tissue engineering skin to repair deep burn wounds.