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

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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