Tissue Engineered Neoesophagus

1.大鼠食管角蛋白细胞的分离和培养1)取供体大鼠的食道黏膜。它们在消解酶溶液中孵育，然后在胰酶溶液中搅拌。成功地修复了大鼠的食道角质形成细胞。2)用3T3饲养层方法在胶原凝胶上成功地培养了食管角化细胞。2)将食道角质形成细胞片植入大鼠背阔肌或背阔肌，并再生了新的食道。1)将切开的角质形成细胞胶原片植入受体大鼠的背阔肌或大网膜，并卷曲缝合。植入后1周、2周、3周取材进行组织学检查。肉眼和显微镜下均证实它们形成了管状组织和新的食道。新食道与小肠吻合术及颈段食管代食管术1)新食道男性大网膜与自然空肠端行…吻合更多的是为了终结时尚。这些大鼠还接受了Roux-Y手术，吻合口不进入食物通道。手术大鼠全部成活，吻合成功。2)背阔肌制作的新食道在植入时被拉至颈部。移植后1周，切除受体大鼠颈段1 cm长的食道，代之以新的食道，端端吻合。术后6d存活，吻合成功。人食道角质形成细胞培养及裸鼠移植1)取食道癌患者的正常食道黏膜片，按上述方法获取角质形成细胞。2)将胶原片植入裸鼠背阔肌上。16天时，附壁细胞层与正常人的食道上皮相似。较少

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