Tissue Engineered Neoesophagus

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1.Isolalon and culture of rat's esophageal keratinocytes1) Donor rats were sacrifioed and pieces of esophageal mucosa were obtained. They were incubated in dispase solution and then stirred in trypsin solution. Rat's esophageal keratinocytes were successfully harvested.2) Esophageal keratiocyies were successfully cultured by 3T3 feeder method on collagen gel.2.Implantation of the esophageal keratinocytes sheet to the omentun or to the latissimus dorsi muscle and regeneration of neoesophagus1) The collagen sheets with cuttured keratinocytes were implanted in recipient rat's latissimus dorsi muscle or in omentun and rolled and sutured. One 2, 3, week after implantation the tissue were taken and examined. They formed tube and neoesophagi were confirmed to regenerate macroscopically and microscopically.3. Anastomosis between Neoesophagus and small intestine and replacement of the cervical esophagus with neoesophagus1)Neoesophagus male in omentum were anastomosed to native jejunum in an end … More to end fashion. These rats also received Roux-Y operation and anastomoses were kept from food passage. Operated rats were all survived and the anastomoses were successfully made. Mucosa of neoesophagi were maintained and microscopically grew and became simliar to normal esophageal mucosa.2) Neoesophagus made in latissimus dorsi muscle was pulled up to neck at the time of implantation. One week after implantation the 1cm cervical esophagus of recipient rat were resected and replaced with neoesophagus and anastomosed them an end to end fashion. The rats survived 6 days after the operation and anastomoses were successfully made.4. Human esophageal keratinocyte culture and implantation to nude mice1) Normal human esophageal pieces of mucosa were obtained from esophageal cancer patients and keratinocytes were harvested as above method. They were cultured on collagen gel with serum free medium.2) The collagen sheets were implanted on latissimus dorsi muscle of nude mice. By 16 days the epithetial cel layer was similar to normal human esophageal epithetum. Less