Investigating endothelial invasion and sprouting behavior in three-dimensional collagen matrices
- 3 December 2009
- journal article
- Published by Springer Science and Business Media LLC in Nature Protocols
- Vol. 4 (12), 1888-1898
- https://doi.org/10.1038/nprot.2009.221
Abstract
Seeding a monolayer of primary human endothelial cells on the surface of a polymerized three-dimensional collagen matrix in the presence of pro-angiogenic stimuli allows manipulation and analysis of rapid sprouting responses. This protocol is useful for elucidating incompletely defined intracellular mechanisms downstream of pro-angiogenic factors that regulate sprout formation and initiation, and can also be used to test the efficacy of pro-and anti-angiogenic compounds. We present protocols to culture endothelial cells, prepare three-dimensional collagen matrices and quantify and image rapid endothelial sprouting responses (24 h). This protocol can be carried out using either type I or type II collagen matrices with primary endothelial cells isolated from macrovascular and microvascular sources of varying species.Keywords
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