An in vitro model of angiogenesis: basic features.
An in vitro model of angiogenesis: basic features.
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DOI：
10.1023/a:1026546219962
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发表时间：
1999-01-01
期刊：
影响因子：
9.8
通讯作者：
Wheatley, D N
中科院分区：
文献类型：
--
作者：
Bishop, E T;Bell, G T;Wheatley, D N
本报告描述了一种血管生成模型，该模型在人脐静脉内皮细胞（HUVEC）和成人患者皮肤来源的人二倍体成纤维细胞的混合物（共培养物）中发展。该系统不需要添加除内皮生长培养基（EGM）中正常存在的生长因子以外的其他生长因子，也不需要添加基质蛋白，并且允许在标准低（2%）浓度的胎牛血清中发生细胞生长和增殖。血管生成是专门刺激血管内皮生长因子（VEGF），导致类似微血管床的结构的发展增加。或者，通过加入过量的中和抗VEGF抗体和抗血管生成药物如苏拉明来抑制血管生成。我们简要地表明，刺激和抑制活动可以很容易地和快速量化的图像分析。通过共聚焦显微镜和电子显微镜证实了小管形成，并对共培养物中这些结构的发育和分布进行了免疫化学分析，以显示特定内皮细胞决定簇（如PECAM-1）的表达。在这一点和其他一些标准上，研究结果验证了这种体外过程作为体内血管生成的模型，可以量化以测定刺激和抑制剂，信号和药物。
This report describes a model of angiogenesis which develops in admixtures (co-cultures) of human umbilical vein endothelial cells (HUVEC) and human diploid fibroblasts of dermal origin from adult patients. The system does not require the addition of further growth factors other than those normally present in endothelial growth medium (EGM), nor matrix proteins, and cell growth and proliferation are allowed to occur in a standard low (2%) concentration of fetal calf serum. Angiogenesis was specifically stimulated in response to vascular endothelial growth factor (VEGF), resulting in an increased development of structures resembling a microvasculature bed. Alternatively, angiogenesis was inhibited by addition of an excess of neutralising anti-VEGF antibodies, and the anti-angiogenic drugs such as suramin. We briefly show that stimulatory and inhibitory activities can be easily and quickly quantified by image analysis. Tubule formation was confirmed by confocal and electron microscopy, and the development and disposition of these structures within the co-cultures has been analysed immunochemically to show expression of specific endothelial cell determinants, such as PECAM-1. On this and a number of other criteria, the findings validate this in vitro process as a model of in vivo angiogenesis that can be quantified to assay stimulatory and inhibitory agents, signals and drugs.