The thin prep rat aortic ring assay: A modified method for the characterization of angiogenesis in whole mounts

被引:54
作者
Zhu W.-H. [1 ]
Nicosia R.F. [1 ,2 ]
机构
[1] Department of Pathology, University of Washington, Seattle, WA
[2] Division of Pathology, VA Puget Sound Health Care System, Seattle
基金
美国国家科学基金会;
关键词
Angiogenesis; Collagen; Confocal microscopy; Endothelium; Rat aorta model; Whole mount;
D O I
10.1023/A:1021509004829
中图分类号
学科分类号
摘要
The rat aortic ring model has gained broad acceptance as an angiogenic assay. This system can be used to study the activity of angiogenic and anti-angiogenic factors, and investigate the molecular mechanisms of the angiogenic process. We describe here a thin prep modification of the aortic ring model, which significantly simplifies the procedure and allows staining of aortic outgrowths as whole mounts. Using this procedure, intact preparations of angiogenic outgrowths are successfully and reproducibly stained with endothelial cell (anti-CD-31 and -Tie2 antibodies, Griffonia Simplicifolia isolectin-B4) and smooth muscle cell (anti-α-smooth muscle actin antibody) markers. Combined use of double immunostaining and confocal microscopy allows concurrent visualization of endothelial and mural cells in the same cultures. Whole mount immunostains of rat aorta cultures are an effective way to rapidly characterize the cellular composition of the angiogenic outgrowths, and localize proteins implicated in the regulation of angiogenesis. This method should facilitate the work of the many vascular biologists that have adopted the rat aorta model as a tool to study angiogenesis and its mechanisms.
引用
收藏
页码:81 / 86
页数:5
相关论文
共 28 条
[1]  
Folkman J., Angiogenesis in cancer, vascular, rheumatoid and other disease, Nat Med, 1, pp. 27-31, (1995)
[2]  
Ferrara N., Alitalo K., Clinical applications of angiogenic growth factors and their inhibitors, Nat Med, 5, pp. 1359-1364, (1999)
[3]  
Nelson M.A., Passeri J., Frishman W.H., Therapeutic angiogenesis: A new treatment modality for ischemic heart disease, Heart Dis, 2, pp. 314-325, (2000)
[4]  
Folkman J., Tumor angiogenesis: Therapeutic implications, N Engl J Med, 285, pp. 1182-1186, (1971)
[5]  
Boehm T., Folkman J., Browder T., Et al., Antiangionenic therapy of experimental cancer does not induce acquired drug resistance, Nature, 390, pp. 404-407, (1997)
[6]  
Kim K.J., Li B., Winer J., Et al., Inhibition of vascular endothelial growth factor-induced angiogenesis suppresses tumour growth in vivo, Nature, 362, pp. 841-844, (1993)
[7]  
Saaristo A., Karpanen T., Alitalo K., Mechanisms of angiogenesis and their use in the inhibition of tumor growth and metastasis, Oncogene, 19, pp. 6122-6129, (2000)
[8]  
Bicknell R., Vascular targeting and the inhibition of angiogenesis, Ann Oncol, 5, SUPPL. 4, pp. 45-50, (1994)
[9]  
Billington D.C., Angiogenesis and its inhibition: Potential new therapies in oncology and non-neoplastic diseases, Drug Des Discov, 8, pp. 3-35, (1991)
[10]  
Aiello L.P., Pierce E.A., Foley E.D., Et al., Suppression of retinal neovascularization in vivo by inhibition of vascular endothelial growth factor (VEGF) using soluble VEGF-receptor chimeric proteins, Proc Natl Acad Sci USA, 92, pp. 10457-10461, (1995)