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目的培养人视网膜微血管内皮细胞,建立人视网膜血管体外二维模型。方法人视网膜血管内皮细胞在纤维连接蛋白包被的细胞培养池内,以无血清人内皮细胞培养基培养,形成二维血管模型。用辣根过氧化酶检测其通透性。部分血管模型加入5ng/ml的血管内皮生长因子培养,与无血管内皮生长因子培养形成的血管模型比较通透性的变化,观察血管内皮生长因子对血管通透性的影响。结果2～4d左右内皮细胞亚融合形成网状血管样结构,6d左右形成较完整的二维血管模型。血管内皮生长因子可增大血管的通透性,促进血管生成。结论用人内皮细胞培养基可成功培养人视网膜血管内皮细胞;利用细胞培养池和无血清人内皮细胞培养基培养,可建立标准的体外视网膜二维血管模型。
Objective To culture human retinal microvascular endothelial cells and establish a two-dimensional model of human retinal blood vessels in vitro. Methods Human retinal vascular endothelial cells were cultured in serum-free human endothelial cell culture medium in fibronectin-coated cell culture to form a two-dimensional vascular model. Permeability of horseradish peroxidase was measured. Vascular endothelial growth factor (VEGF) was cultured in 5ng / ml vascular models in part of the vascular models. The changes of permeability were compared with those of the vascular models without vascular endothelial growth factor (VEGF) to observe the effects of vascular endothelial growth factor on vascular permeability. Results The endothelial cells sub-fusion formed reticular vascular structures around 2 ~ 4 days, forming a more complete two-dimensional vascular model around 6 days. Vascular endothelial growth factor increases vascular permeability and promotes angiogenesis. Conclusion Human retinal vascular endothelial cells can be successfully cultured with human endothelial cell culture medium. A standard in vitro two-dimensional retinal vascular model can be established by culturing with cell culture pool and serum-free human endothelial cell culture medium.