Investigating endothelial invasion and sprouting behavior in three-dimensional collagen matrices.
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.
Bayless
Kayla J
KJ
Department of Molecular & Cellular Medicine, Texas A&M Health Science Center, College Station, Texas, USA. kbayless@medicine.tamhsc.edu
Kwak
Hyeong-Il
HI
Su
Shih-Chi
SC
D043925:Angiogenesis Inducing Agents / Q000494:pharmacology; D000818:Animals; D018929:Cell Culture Techniques; D003094:Collagen; D042783:Endothelial Cells / Q000502:physiology*; D004730:Endothelium, Vascular / Q000502:physiology; D049452:Green Fluorescent Proteins / Q000032:analysis; D006801:Humans; D018919:Neovascularization, Physiologic; D051381:Rats
D043925:Angiogenesis Inducing Agents; D049452:Green Fluorescent Proteins; D003094:Collagen