碱性成纤维细胞生长因子对三维培养内皮细胞血管样结构形成的作用研究

Effect of Basic Fibroblast Growth Factor on Formation of Vascular Tube in Three-Dimensional Cell Culture

目的 :观察碱性成纤维细胞生长因子 (basicfibroblastgrowthfactor,bFGF)对三维培养内皮细胞血管样结构形成的作用。方法 :实验分为两组 ,即加入浓度 2 0ng·ml-1bFGF的bFGF组及不加bFGF的对照组。采用Pepper等介绍的三维培养胶原基质配制及细胞培养方法制作标本 ,透射电子显微镜下观察结果。 结果 :对照组内皮细胞在胶的表面呈单层生长 ,未见其向深层生长及管腔结构形成。bFGF组内皮细胞在胶内呈多层浸润生长 ,伸展、变形呈扁平状 ;内皮细胞与胶原纤维之间可见半桥粒结构 ,内皮细胞之间以桥粒连接 ,形成毛细血管样管腔结构 ;在形成管状结构的内皮细胞中可见空泡样变 ,边界由胞质与胞膜组成 ,细胞核变形 ,凹陷 ,可见切迹 ,或呈薄片状 ;内皮细胞游离面有微绒毛突起。结论 :三维培养技术简单易行 ,是揭示在体血管形成的最简单模型 ;bFGF有利于三维培养内皮细胞在有展性的胶原基质面形成血管样结构。

Objective: To observe effect of basic fibroblast growth factor (bFGF) on the formulation of tubular structures in a three-dimensional cell culture system. Methods: Endothelial cells were cultured in a three-dimensional system on collagen gel surface through the stimulation of bFGF. Results: In bFGF group, endothelial cells grew in multi-layer soak type and transmuted into flat cells. Endothelial cells connected to collagen through hemidesmosome and themselves connected with each other through desmosome to form vascular tubes. Vascular tubes had various sizes and there were many endothelia cells in them. Endothelial cells and their nucleus changed in shape. In control group, endothelial cells grew in single layer and did not form vascular tubes. Conclusion: The technique of three-dimensional cell culture is simple and practicable for observation of angiogenesis in vivo. It is found that bFGF in three-dimensional cell culture is helpful for forming tubular structures.