Development of a Hybrid Artificial Esophagus Using Cultured Human Esophageal Epithelial cells, fibloblasts, smooth muscle cells

1.利用成纤维细胞促进人工食管壁上皮细胞的增殖和分化。方法：将食管成纤维细胞包埋在直径为35 mm的培养皿中，用聚乙醇酸网片包埋在I型胶原溶液中。将食管上皮细胞接种于胶原片上，与KGM共同培养，待细胞融合后，缝合PGA网片的两端，制成黏膜管。将无胸腺大鼠背部切开，制作背阔肌肌瓣。用纤维胶将粘膜管包裹在肌瓣中。移植后第14天取出移植的粘膜管，进行上皮细胞分层检查。结果：术后14天取出的管腔结构无狭窄，分层20层。上皮细胞的分化程度从基底层到最上层呈递增的梯度 ...更多信息 离子也被观察到。胶原层内可见肌层血管生成。2.将胶原片包埋于成纤维细胞胶原层外，观察平滑肌细胞能否包绕在人工食管壁周围作为肌层。方法：将人主动脉平滑肌细胞包埋在I型胶原中。然后将人真皮成纤维细胞包埋在具有平滑肌细胞的上述胶原层上的I型胶原中。最后，将人食管上皮细胞培养在成纤维细胞的胶原层上。将构建的胶原膜移植于裸大鼠背阔肌上。在1周和2周后，用苏木精-伊红染色法和免疫染色法（抗人肌动蛋白抗体：HHF 35）对薄片进行组织学检查。结果如下：1周后取出的胶原片的组织学特征如下：（1）平滑肌细胞层位于胸壁下层，与成纤维细胞层有明显区别;（2）平滑肌细胞沿胸壁弯曲方向呈低密度层间排列;两周后取出的组织学特征如下：1.平滑肌细胞沿弯曲方向排列，细胞密度较高; 2.平滑肌细胞和成纤维细胞均可见血管生成; 3.上皮细胞分层10- 13层。上皮层、粘膜下层和固有肌层均得到重建。少

1.We utilized fibroblasts to acelerate proliferation and differentiation of epithelial cells of the artificial esophageal wall. Methods : Esophageal fibroblasts were embedded in type I collagen solution with polyglycolic acid mesh in a 35mm diameter culture dish. Esophageal epithelial cells were seeded on the collagen sheets and cultured with KGM.When the culture cells reached confluency, both edges of the PGA mesh were sutured to make a mucosal tube. The backs of athymic rats were incised and muscle flaps in the latissimus dorsi muscle were made. The mucosal tubes were wrapped in the muscle flaps using fibrine glue. Grafted mucosal tubes were removed on the 14th day after transplantation and epithelial cell stratification examined. Results : Luminal structures removed on the 14th day also did not have stenosis, and had twenty layr stratification. Numerous flat epithelial cells in an increasing gradient from the basal cells to the uppermost layr were found. Epithelial cell differentiat … More ion was also observed. Angiogenesis from the muscle layr was observed in the collagen layr.2.We examined whether smooth muscle cells could be surrounded around the artificial esophageal wall as a muscle layr by laminificating collagen sheet embedding smooth muscle cells outside of the collagen layr of fibroblasts. Methods : Human aortic smooth muscle cells were embedded in type I collagen. Human fibroblasts from dermis were then embedded in type I collagen on the abovementioned collagen layr with smooth muscle cells. Finally, human esophageal epithelial cells were cultured on the collagen layr of the fibroblasts. The constructed collagen sheet were transplantated on the lattisimus dorsi muscles of nude rats. The sheets were examined histologically at one and two weeks later under hematoxillin-eosin stain and immunological stain methods (anti human actin antibody : HHF 35). Results : The histological features of the collagen sheet extirpated one week later were as follows : 1.The layr of smooth muscle cells was found in the lower layr and was clearly distinguishable from the layr of fibroblasts, 2.Smooth muscle cells arrayd layr to layr in a low cell density in a direction of curvature of the chest wall, 3.Epithelial cells stratified 6-8 layrs. The histological features of the sheet extirpated two weeks later were as follows : 1.Smooth muscle cells arrayd in a high cell density in a direction of the curvature, 2.Angiogenesis was found in both layrs of the smooth muscle cells and fibroblasts, 3.Epithelial cells stratified 10-13layrs. It seemed as if the epithelial layr, submucosal tissue layr and proper muscle layr were reconstructed. Less