Abstract
Angiogenesis is a multifactorial event which requires the migration, proliferation, differentiation and structure rearrangement of endothelial cells. This angiogenic process has been commonly studied using in vitro assays such as Boyden chamber assay, wound healing assay and tube formation assay. These assays mainly use monolayers of endothelial cells which are modified by repeated passages and are fully proliferative, a situation far away from physiology. In addition, not only endothelial cells are involved in this process but surrounding cells (such as pericytes, smooth muscle cells, fibroblasts) and the supporting matrix are also major players. The three-dimensional ex vivo aortic ring model recapitulates the complexities of angiogenesis and combines the advantages of in vitro and in vivo models. The aortic ring is cultivated in a chemically defined culture environment. Microvessels which grow in this system are lumenized vessels with surrounding supporting cells and are essentially indistinguishable from microvessels formed during angiogenesis in vivo. The efficacy of pro-or anti-angiogenic factors can be determined in the absence of serum molecules which may otherwise interfere with the substances being tested (Nicosia and Ottinetti, 1990). However, this system requires access to fresh rat tissue but several samples can be prepared from one aorta.
Keywords: Aortic ring, Angiogenesis, Endothelial cells, Ex vivo model
Materials and Reagents
Equipment
Procedure
The day before Wrapped dissecting board in 2 aluminium foils Sterilize: Beaker, magnetic bar, blades and dissecting materials Put in a fridge: 200 µl tips box, Cell culture plates 48 well On the day
Representative data
Figure 1. Representative pictures of one aortic ring at different times in culture (top left to bottom left) and pictures of aortic rings treated with a growing concentration of an angiogenic agent (from left to right) at 8 days post-isolation
Notes
Recipes
Acknowledgments
This protocol was adapted from Nicosia and Ottinetti (1990) and Blacher et al. (2001). This study was supported by the Society for Research on Cardiovascular Diseases and The Ministry of Culture, Higher Education and Research of Luxembourg.
References
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