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 m eans to simplify, accelerate and improve the efficiency with which prolifer ating cells are delivered to full thickness or non-healing skin defects. Ho wever. there have been no studies that monitor the transfer of cells from s uch membranes to the wound bed. We have used a porcine model of lacZ gene m arked cultured autologous keratinocyte grafting to demonstrate unambiguousl y the transfer of cultured cells to cutaneous wounds from the EpiGen(TM) po lymer membrane developed by Smith & Nephew Group pie. Full thickness wounds enclosed within rigid chambers were first grafted with autologous de-epide rmalised dermis (DED). Keratinocytes were cultured on EpiGen(TM) membranes and applied to the wound beds 7 days after the DED grafts. Epidermal remnan ts persist within the DED and the resultant epidermis is therefore, a mixtu re of wound regeneration and delivered cultured cells. Unequivocal evidence for keratinocyte transfer from the membrane was obtained through the obser ved macroscopic surface staining for lacZ transduced cells and lacZ positiv e cells detected in sections through deeper layers of epidermal tissue. Thi s method offers a general approach for evaluating the efficiency of keratin ocyte delivery using upside-down flexible membrane transfer. (C) 2001 Elsev ier Science Ltd and ISBI. All rights reserved.