The combination of cultured autologous keratinocytes with the dermal regeneration template Integra™ could offer increased possibilities for reconstructive surgery and wound healing. A single-step application of cells, centrifuged deep into an Integra™-like matrix at the silicone–matrix junction, has been described but might prove technically complex for clinical use. We have investigated the possibility of simplifying this procedure by applying cultured cells directly to the underside of the Integra™ or directly to the wound bed immediately prior to grafting. The objective was to see whether cells would migrate through the matrix in an upward direction. We tested the principle of this concept using a pig wound healing model. Integra™ was seeded directly with cultured cells and grafted onto fresh full-thickness wounds, or unseeded Integra™ was applied to freshly excised wound beds that had just been seeded with the same number of cells. Biopsies were taken at 3, 7, 11, and 14 days. Histological sections showed that the cells moved through the Integra™ to give a confluent surface epithelium. Direct seeding onto the Integra™ was the most efficient method. Transduction of cultured autologous keratinocytes in vitro with a MFGlacZnls retrovirus confirmed that the epidermis was derived from the cultured autologous keratinocytes.