Polyvinyl alcohol (PVA) hydrogel and stem cell therapy have been widely used in wound healing. However, the lack of bioactivity for PVA and security of stem therapy limited their application. In this study, an adipose-derived stem cells (ADSCs)-seeded PVA dressing (ADSCs/PVA) was prepared for wound healing. One side of the PVA dressing was modified with photo-reactive gelatin (Az-Gel) via ultraviolet (UV) irradiation (Az-Gel@PVA), and thus ADSCs could adhere, proliferate on the PVA dressings and keep the other side of the dressings without adhering to the wound. The structure and mechanics of Az-Gel@PVA were determined by scanning electron microscopy (SEM) and material testing instruments. Then, the adhesion and proliferation of ADSCs were observed via cell counts and live-dead staining. Finally, in vitro and in vivo experiments were utilized to confirm the effect of ADSCs/PVA dressing for wound healing. The results showed that Az-Gel was immobilized on the PVA and showed little effect on the mechanical properties of PVA hydrogels. The surface-modified PVA could facilitate ADSCs adhesion and proliferation. Protein released tests indicated that the bioactive factors secreted from ADSCs could penetrated to the wound. Finally, in vitro and in vivo experiments both suggested the ADSCs/PVA could promote the wound healing via secreting bioactive factors from ADSCs. It was speculated that the ADSCs/PVA dressing could not only promote the wound healing, but also provide a new way for the safe application of stem cells, which would be of great potential for skin tissue engineering.