Improved quantification of angiogenesis in the rat aortic ring assay

被引:84
作者
Blacher S. [1 ]
Devy L. [1 ]
Burbridge M.F. [2 ]
Roland G. [1 ]
Tucker G. [2 ]
Noël A. [1 ]
Foidart J.-M. [1 ]
机构
[1] Laboratory of Tumor and Developmental Biology, University of Liège, CHU, Sart-Tilman, Liège
[2] Experimental Oncology Division, Institut de Recherches Servier, Suresnes
关键词
Angiogenesis; Image analysis; Protease inhibitors; Rat aortic ring model; VEGF;
D O I
10.1023/A:1012251229631
中图分类号
学科分类号
摘要
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.
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页码:133 / 142
页数:9
相关论文
共 24 条
[1]  
Folkman J., Brem H., Angiogenesis and inflammation, Inflammation: Basic Principles and Clinical Correlates, pp. 809-839, (1996)
[2]  
Folkman J., Angiogenesis in cancer, vascular, rheumatoid and other disease, Nature Med, 1, pp. 27-31, (1995)
[3]  
Folkman J., Klagsbrun M., Angiogenic factors, Science, 235, pp. 442-447, (1987)
[4]  
Auerbach R., Auerbach W., Polakowski I., Assays of angiogenesis: A review, Pharmacol Ther, 51, pp. 1-11, (1991)
[5]  
Nicosia R.F., Ottinetti A., Growth of microvessels in serum-free matrix culture of rat aorta: A quantitative assay of angiogenesis in vitro, Lab Invest, 63, pp. 115-122, (1990)
[6]  
Kobayashi S., Fukuta M., Kontani H., Et al., A quantitative assay for angiogenesis of cultured choroidal tissues in streptozotocin-diabetic wistar and spontaneously diabetic GK rats, Jpn J Pharmacol, 78, pp. 471-478, (1998)
[7]  
Brown K.J., Maynes S.F., Bezos A., Et al., A novel in vitro assay for human angiogenesis, Lab Invest, 75, pp. 539-555, (1996)
[8]  
Bocci G., Danesi R., Benelli U., Et al., Inhibitory effect of suramin in rat models of angiogenesis in vitro and in vivo, Cancer Chemother Pharmacol, 43, pp. 205-212, (1999)
[9]  
Kruger E.A., Duray P.H., Tkosos M.G., Et al., Endostatic inhibits microvessel formation in the ex vivo rat aortic ring angiogenesis assay, Biochem Biophys Res Commun, 268, pp. 183-191, (2000)
[10]  
Nissanov J., Tuman R.W., Gruver L.M., Fortunato J.M., Automatic vessel segmentation and quantification of the rat aortic ring assay of angiogenesis, Lab Invest, 73, pp. 734-739, (1995)