The study of an artificial esophagus constructed of cultured human esophageal wall cells with angiogenetic effects

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We have been developing artificial esophagus which is constructed of autologous cells proliferated by the cell culture methods and extracellular matrix. In a previous study by Sato and Miki, our co-culture methods of esophageal epithelial cells and fibroblasts had been established1. We tried to consturct an artificial csophagus which consisted of human csophageal epithelial cells, dermal fibroblasts, smooth muscle cells isolated from aortic media with the aim of substituting the elasticity and the support of the csophageal wall. 15X105 cells of smooth muscle cells were embedded in 1.5 ml of type I collagen 6X105 cells of human fibroblasts were then embedded in 0.7 ml of type I collagen on the above-described collagen layer with smooth muscle cells. Finally, human esophagcal epithelial cells were cultured on the collagen layer of the fibroblasts. The constructed collagen sheets were cultured in vitro for one week, then transplanted on the lattisimus dorsi muscle of athymic rats. The sheets were examined histologically at one and two weeks later under hematoxillin-eosin stain and immunologicaly stained methods (anti actin antibody). One week later, epithelial cells stratified and the lower layer of smooth muscle cells which arrayed in a low cell density toward the direction of curvature of the chest wall were distinguishable from the upper layer of fibroblasts. And the angiogencsis was recognized in the lower layer. Two weeks later, the collagen sheet seemed microscopically as if the epithelial layer, the submucosal layer and the proper muscle layer were reconstructed.Additionaly, we succeeded in isolating of the intestinal smooth musclecells from the colon of a mice. If the collagen sheet consisted of esophageal epithelial cells, fibroblasts and smooth muscle cells from the surgically resected specimen can be transformed to the luminal structure, possible clinical application of the artificial esophagus is suggested.