Integra is a skin substitute used for dermal reconstruction. Current clinical practice consists of two procedures, first applying Integra to the wound and then replacing the silicone pseudo-epidermis with an epidermal autograft 3 weeks later. This two-step repair limits the clinical use of the product. An effective one-step procedure could reduce the time taken to repair and decrease the number of procedures for use of Integra. This study examined the effects of simultaneous application of a non-cultured autologous suspension of cells, isolated using the ReCell autologous cell harvesting device, in combination with Integra, to achieve a one-step skin repair. In two female Yorkshire swine, 10 full-thickness wounds were created. Wounds were treated with Integra seeded with cell suspension and compared to controls of Integra alone and cell suspension alone. Weekly macroscopic and histological assessment demonstrated that the wounds treated simultaneously with Integra and non-cultured autologous cells had enhanced epithelialization at an early time-point compared to controls. Wounds treated simultaneously with Integra and cell suspension demonstrate that cells remain viable, migrate through the Integra template and self-organize into differentiated epidermis. The results indicate that combining Integra with autologous cells facilitates one-step skin reconstruction of a full-thickness skin wound.