A novel culturing and grafting system for the treatment of leg ulcers

The purpose of this study was to develop and test an efficient culturing and grafting system for the treatment of leg ulcers. The culturing system consisted of a Petriperm culture vessel (20 cm2) aseptically placed in a larger standard Petri dish (60 cm2). Skin cultures were established and cultivated in the Petriperm dish. The cells grew on the bottom of the Petriperm dish, which was made of a gas-permeable 25-micron thick transparent Teflon film. Grafts were produced simply by cutting the film from the bottom of the Petriperm dish with a scalpel. The system was used to produce subconfluent epidermal autografts which were used to heal a 32 cm2 chronic rheumatoid arthritis leg ulcer. The cultured autografts were transferred cell side down on to the cleaned wound bed without an enzymatic digestion. The grafts consisted of autologous keratinocytes, melanocytes and fibroblasts. Caution was taken not to disturb the wound bed for 7-9 days at which time the Teflon film was removed. The wound closed 2 weeks after the last grafting and has remained closed for more than a year post-treatment. The culturing and grafting system presented here will make it possible to develop cellular-based therapies that were previously not possible.