关键词: 角膜上皮, 角膜缘干细胞, 纤维蛋白

Abstract: Abstract:Objective To reconstruct the fibrin-based tissue engineering corneal epithelium using limbal stem cells as seed cells and provide new grafting material for corneal epithelium transplantation.Methods The fibrin-based frame was reconstructed by mixing 20 m•L^-1 fibrinogen solution and 10 U•mL^-1 lthrombin and the normal struture and ultrastructure of frame was detected. The limbus corneal tissue (2 mm×2 mm)was obtained,the fibrin-based frame after trypsind egestion.The growth status of the cells was obsewed ,and the changes of morphology,ultrastructure and immunohistochemistry were observed after two weeks. Results Fibrin-based frame was soft and had extensibility.Its dimension was 70-108 μm,the average empty porosity was 70.4%.It was multilayer network observed under scanning electron microscope.The cells migrated from the edge of the tissue 36 h after inoculation,single layer cells were formed after 7 d, the cells were combined with fibrin to form gragting implant after 14 d.The cells remained the typical ultrastructure feature of the epithelium observed under transmission electron microscope.The immunohistochemical results of the anti- cytokeratin McAb AE5 and the anti-p63 McAb 4A4 were positive. Conclusion Fibrin is a suitable frame material for corneal epithelium transplantation,provided a new grafting material for tissue engineering corneal epithelium.

Key words: corneal epithelium, limbal stem cells, fibrin