兔角膜上皮-基质层组织工程方法三维重建

Reconstructed corneal epithelium-stroma producted in vitro by tissue engineering

  • 摘要: 目的: 利用细胞生物学和工程学原理在体外重建角膜上皮和基质层,提供与正常生理结构相似的模型。方法: 兔角膜上皮和基质细胞原代并传代培养,免疫组化方法对细胞进行鉴定,以鼠尾为原料提取I型胶原溶液,重建角膜的基质和上皮,上皮实行气液界面培养,倒置显微镜观察细胞形态。36h、72h、7d、14d、2 0d重建组织常规病理切片。结果: 倒置显微镜下角膜成纤维细胞呈梭形三维生长,上皮细胞多层生长。组织病理检查显示重建的角膜组织结构类似于天然角膜组织。结论: 本实验可为角膜的病理生理、毒理、药理以及基因表达等研究提供有利的模型和工具。

     

    Abstract: Objective: To reconstruct corneal tissue with epithelium and stroma by tissue engineering.Methods: Epithelial cells and keratocytes were isolated from rabbit corneas and cultured on plastic substrates in vitro They could be subcultured for several passages Using a special transwell, we succeeded to culture two kinds of cells without mixture each other Anti vementin and anti keratin antibodies were used to identify the cells Reconstructed corneal tissues were obtained by culturing epithelial cells on collagen gels containing keratocytes The topmost layer, Consisting of corneal epithelial cells was treated by what was designed as the air liquid interface method The tissue was prepared for histological at 36 h,72 h,7 d,14 d and 20 d.Results: The corneal epithelium and keratocytes cultured in the collagen gel were different from those cultured on plastic morphologically and biochemically, which showed appropriate histological structure Conclusions: The tissue engineered cornea in vitro provides ideal toolsfor further physiological, toxicological and pharmacological studies as well as being an attractive model for gene expression studies

     

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