Demonstration of epidermal transfer from a polymer membrane using genetically marked porcine keratinocytes

The culture of keratinocytes on flexible membranes has been proposed as a means to simplify, accelerate and improve the efficiency with which proliferating cells are delivered to full thickness or non-healing skin defects. However, there have been no studies that monitor the transfer of cells from such membranes to the wound bed. We have used a porcine model of lacZ gene marked cultured autologous keratinocyte grafting to demonstrate unambiguously the transfer of cultured cells to cutaneous wounds from the EpiGen polymer membrane developed by Smith & Nephew Group plc. Full thickness wounds enclosed within rigid chambers were first grafted with autologous de-epidermalised dermis (DED). Keratinocytes were cultured on EpiGen membranes and applied to the wound beds 7 days after the DED grafts. Epidermal remnants persist within the DED and the resultant epidermis is therefore, a mixture of wound regeneration and delivered cultured cells. Unequivocal evidence for keratinocyte transfer from the membrane was obtained through the observed macroscopic surface staining for lacZ transduced cells and lacZ positive cells detected in sections through deeper layers of epidermal tissue. This method offers a general approach for evaluating the efficiency of keratinocyte delivery using upside-down flexible membrane transfer.