Improved quantification of angiogenesis in the rat aortic ring assay
Improved quantification of angiogenesis in the rat aortic ring assay
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DOI：
10.1023/a:1012251229631
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发表时间：
2001-01-01
期刊：
影响因子：
9.8
通讯作者：
Foidart, Jean-Michel
中科院分区：
文献类型：
--
作者：
Blacher, Silvia;Devy, Laetitia;Foidart, Jean-Michel
体外血管生成试验对于鉴定潜在的血管生成剂和筛选药理学抑制剂是必不可少的。在这些实验中，Nicosia开发的大鼠主动脉环模型填补了体内和体外模型之间的空白。该系统上血管生成的量化必须适用于表征各种复杂状态的血管网络。我们在这里提出了一种改进的计算机辅助图像分析方法，它允许：(1)确定主动脉环面积及其因子形状；(2)微血管数量、分支总数、最大微血管长度和微血管分布；(3)分离成纤维细胞样细胞总数及其分布。我们表明，这种方法适用于量化自发血管生成，以及分析由不同浓度的血管内皮生长因子（VEGF）诱导的复杂微血管网络。此外，通过评估一个新的参数，即成纤维细胞样细胞分布，我们的结果表明：(1)在自发血管生成反应中，最大的成纤维细胞样细胞迁移界限微血管的生长；(2)已知的血管生成抑制剂Batimastat可以阻止内皮细胞发芽，但不会完全阻断成纤维细胞样细胞的迁移。最后，这种新的定量方法对更好地了解血管生成和测试促血管生成或抗血管生成药物有很大的兴趣。
In vitro angiogenesis assays are essential for the identification of potential angiogenic agents and screening for pharmacological inhibitors. Among these assays, the rat aortic ring model developed by Nicosia bridges the gap between in vivo and in vitro models. The quantification of angiogenesis on this system must be applicable to characterise vascular networks of various states of complexity. We present here an improved computer-assisted image analysis which allows: (1) the determination of the aortic ring area and its factor shape; (2) the number of microvessels, the total number of branchings, the maximal microvessel length and the microvessel distribution; (3) the total number of isolated fibroblast-like cells and their distribution. We show that this method is suitable to quantify spontaneous angiogenesis as well as to analyse a complex microvascular network induced by various concentrations of vascular endothelial growth factor (VEGF). In addition, by evaluating a new parameter, the fibroblast-like cell distribution, our results show that: (1) during spontaneous angiogenic response, maximal fibroblast-like cell migration delimits microvascular outgrowth; and (2) the known angiogenic inhibitor Batimastat prevents endothelial cell sprouting without completely blocking fibroblast-like cell migration. Finally, this new method of quantification is of great interest to better understand angiogenesis and to test pro- or anti-angiogenic agents.