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

使用培养的人食管上皮细胞进行混合人工食管的实验研究。

基本信息

批准号：
05671085
负责人：
ANDO Nobutoshi
金额：
$ 1.22万
依托单位：
Keio University
依托单位国家：
日本
项目类别：
Grant-in-Aid for General Scientific Research (C)
财政年份：
1993
资助国家：
日本
起止时间：
1993 至 1994
项目状态：
已结题

项目摘要

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.

用细胞培养法构建人工食管。在人工食管的内侧衬上上皮对防止狭窄和加强管腔是很重要的。将正常人上皮细胞培养在包埋有聚乙醇酸（PGA）补片的胶原上。7天后，将PGA网片边缘缝合制成离体管。将由人上皮和胶原-PGA网组成的管移植到无胸腺大鼠背阔肌肌瓣表面。为了促进食管上皮细胞的分层，将它们分别培养在已包埋0、4和8 × 10^5个细胞/ml的人成纤维细胞的胶原片上。3周后行组织学检查。尽管在没有成纤维细胞的胶原蛋白片上的人上皮细胞分层2-3倍，但在包埋有成纤维细胞4*10^5个细胞/ml的胶原蛋白上，它们分层4-5倍。将由人上皮、4 × 10^5细胞/ml成纤维细胞和胶原组成的片移植到无胸腺大鼠的腹直肌表面。2周后行组织学检查。成纤维细胞可促进血管从直肌层向胶原层的生长，并使上皮细胞层化10倍以上。成纤维细胞最终可用于人工食管内侧的分层。