Experimental Study of a Hybrid Artificial Esophagus Using Cultured Human Esophageal Epithelial Cells.

采用细胞培养法重建人工食管。在人工食道内侧铺上上皮细胞以防止狭窄和加强管腔是很重要的。正常人上皮细胞在聚乙醇酸（PGA）网状物包埋的胶原蛋白上培养。7 d后，在体外缝合PGA补片边缘制成试管。将人上皮和胶原- pga网组成的导管移植于胸大鼠背腹肌瓣表面。移植后第20天，管腔结构保持正常，无狭窄。为了加速食管上皮细胞的分层，将它们分别培养在包埋了0、4和8*10^5细胞/ml的人成纤维细胞的胶原片上。3周后进行组织学检查。人上皮细胞在不含成纤维细胞的胶原片上分层2-3倍，在含成纤维细胞4*10^5细胞/ml的胶原片上分层4-5倍。进一步包埋成纤维细胞8*10^5个/ml，成纤维细胞分层17-18倍，分化更大。将人上皮细胞、成纤维细胞（4*10^5个细胞/ml）和胶原蛋白组成的薄片移植到胸腺大鼠腹直肌表面。2周后行组织学检查。观察到从直肌到胶原膜的血管生成和上皮10倍以上的分层。成纤维细胞促进了人上皮细胞的分层。成纤维细胞最终对人工食管内壁的分层非常有用。

An artificial esophagus has been reconstructed by cell culture method. It is important to line inner side of the artificial esophagus with epithelium to prevent strictures and reinforce the lumen. Normal human epithelial cells were cultured on the collagen in which polyglycolic acid (PGA) mesh was embedded. 7 days later, atube was made by suturing the edge of PGA mesh in vitro. The tube consisting of human epithelium and collagen-PGA mesh was transplanted on the surface of alattisimus dorsi muscle flap of an athymic rat. On the 20th day after transplantation, the luminal structure was maintained without strictures.To accelerate stratification of the esophageal epithelial cells, they were cultured on a collagen sheet in which 0,4 and 8*10^5 cells/ml of human fibroblasts were already embedded respectively. They were examined histologically 3 weeks later. Though human epithelial cells on a collagen sheet without fibroblasts stratified 2-3 fold, on collagen in which fibroblasts 4*10^5 cells/ml were embedded, they stratified 4-5 fold. Further fibroblasts 8*10^5 cells/ml were embedded in, they stratified 17-18 fold and differentiated much more.The sheet consisting of human epithelium, fibroblasts 4*10^5 cells/ml and collagen was transplanted on a surface of the abdominal rectus muscle of an athymic rat. It was examined histologically 2 weeks later. Angiogenesis from the rectus muscle into the collagen sheet and more than 10 fold stratification of the epithelium were observed.Stratification of the human epithelial cells were accelerated by the fibroblasts. The fibroblasts are very useful for stratification of inner side of the artificial esophagus eventually.