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目的 :采用ECV30 4细胞 (人脐静脉内皮细胞株 )构建体外血管新生的三维模型 ,为ECV30 4细胞应用于体外血管新生的研究提供依据。方法 :制作鼠尾胶原三维凝胶 ,在凝胶表面种植ECV30 4细胞 ,培养至接近汇合状态 ,换用含碱性成纤维细胞生长因子 (bFGF)的培养液继续培养 3～ 12d ,用倒置相差显微镜观察内皮细胞株向凝胶内迁移、生长状况。结果 :bFGF组ECV30 4细胞迁移到凝胶内不同层次 ,形成细胞条索并吻合成网 ;垂直切面上可见ECV30 4细胞从凝胶表面的单细胞层向凝胶内发芽生长 ,形成树枝状毛细血管样结构。结论 :ECV30 4细胞在三维胶原凝胶基质培养模型中 ,在适当的因子诱导下 ,能够表现出血管新生中的增殖、迁移、发芽等步骤 ,可以作为内皮工具来研究体外血管新生。
OBJECTIVE: To construct a three-dimensional model of ECV304 cells (human umbilical vein endothelial cell line) in vitro and to provide a basis for the study of ECV304 cells in vitro angiogenesis. METHODS: Three-dimensional gel of rat tail collagen was prepared. The ECV30 4 cells were seeded on the surface of the gel. The cells were cultured to confluency and switched to culture medium containing basic fibroblast growth factor (bFGF) for 3 to 12 days. Microscopic observation of endothelial cell migration to the gel, the growth status. Results: The ECV304 cells in bFGF group migrated into different layers of the gel to form cell cords and formed anastomosis network. On the vertical section, ECV30 4 cells germinated and grew into the gel from the single cell layer on the surface of the gel to form dendritic capillary Vascular-like structure. CONCLUSION: ECV304 cells can demonstrate the proliferation, migration and germination in angiogenesis induced by appropriate factors in the three-dimensional collagen gel matrix culture model and can be used as an endothelium to study angiogenesis in vitro.