人工生物可降解支架聚乳酸-聚乙醇酸种植人食管上皮细胞的实验研究

目的研究人食管上皮细胞在聚乳酸-聚乙醇酸(PLGA)三维支架上的贴附和生长情况,利用组织工程技术培养工程化人工食管.方法制作PLGA三维细胞支架;分离培养成人食管上皮细胞,体外扩增后种植到PLGA支架上.在体外和裸鼠体内分别培养食管上皮细胞-支架复合物,分期终止培养,进行组织学染色、扫描电镜、细胞角蛋白免疫组织化学检测.结果体外培养发现,人食管上皮细胞在支架材料上贴附生长良好,长期培养仍保持食管上皮细胞特性,裸鼠体内培养4周后可形成食管黏膜样组织.结论 PLGA支架适合食管上皮细胞黏附生长,可作为食管组织工程的细胞载体.利用组织工程的方法可获得适于移植的工程化人工い食管.

Experimental study on implantation of human esophageal epithelial cells on artificial biodegradable scaffold PLGA

Objective] To verify the adhesion and growth ability of human esophageal epithelial cells(HEECs) on poly(lactic-co-glycolic acid)(PLGA), a three-dimensional biodegradable polymer scaffold, and to reconstruct human esophagus by tissue engineering. Methods] HEECs isolated from adult esophagi resected in esophagectomy were seeded onto PLGA scaffold after expansion by in vitro culture. The complex of cell-scaffold were then cultured ex vivo and transplanted to subcutaneous of nude mouse respectively. The cell-seeded scaffolds in different culture intervals were assessed by histological staining of H-E. Scanning electron microscopy was utilized to verify the interactions between the cells and the biomaterial. Immunohistochemical analyses was also performed to evaluate cytokeratin. Results] HEECs were well distributed and adhered to PLGA scaffolds and maintained their characteristics throughout the culture period. Both the first passaged esophageal epithelial cells and seeded cells could express and product cytokeratin. After cultured in vivo for 4 weeks the cell-seeded scaffolds grew like tissues. Conclusions] PLGA scaffolds can support the esophageal epithelial cells'proliferation, and can be suitable carrier for tissue engineering of artificial esophagus.