Experimental studies on development of the artificial esophagus utilizing cell-seeding method and EGF

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To prevent anastomotic leakages and stricture formations of the artificial esophagus,the inner surface of the prosthesis should be epithelized immediately after implantation. A silicone-coated stainless steal mesh tube was used as the esophageal prothesis,and was implanted under the skin of canine back,wrapping with a prepared latissmus dorsi muscle flap. Two weeks later,the inner surface of the prosthesis was covered with good granulations. As the first trial of epithelizing the inner surface of the prothesis,the skin flap was induced to the inside of the prothesis,and EGF was injected into the prothesis. However thr prothesis suffered intedion frequently and the epithelization was not satisfactory one. Therefore,cell-seeding method was applied. At first,epidermal cells derived enzymatically from the skin were seeded into the lumen of the prosthesis. But epidermal cells were not able to be implanted. The failure of cell-seeding was considered to be attributable to a lack of seeded cells,so cell culture of epidermal cells harvested enzymatically from skin was tried in order to increase cell counts. The method of cell is same as under: 1.Skin was excised. 2.Incubated in DMEM with disease at 37ﾟC for 3 hrs. 3. Epidermis was separated from dermis. 4.Epidermal cells were separated after trypsinzation. 5.Epidermal cells were cultured in KGM. 10 to 14 days later,when epidermal cells were cultured to confluency in flask,trypsinization was done to obtain epidermal cells suspension. Then cultured epidermal cells suspension were seeded into inner surface of the prosthesis. Some of the cultured epidermal cells were able to be implanted on the granulations of the prosthesis. Microscopically,layers of stratified squamous epithelium were seen on the granulations. Based on these results, the epithelization on the inner-surface of the prosthesis is considered to be possible by seeding technique of cultured epidermal cells.