Quantitative and qualitative in vivo angiogenesis assay.
We describe the development and optimization of an in vivo angiogenesis assay utilizing gelfoam sponges impregnated with 0.4% agarose and different proangiogenic factors, such as basic fibroblast growth factor (bFGF), vascular epidermal growth factor (VEGF), tumor growth factor-alpha (TGF-alpha), and endothelial growth factor (EGF). The sponges are implanted into the subcutis of mice and harvested after different times. The gelfoam sponges are fixed, sectioned, and stained with fluorescent antibodies against CD31. The median number of CD31+ cells is determined in 10 different 0.159-mm2 fields. Proangiogenic molecules induced significant migration and proliferation of endothelial cells. To demonstrate the utility of this assay for evaluation of an antiangiogenic agent, mice were implanted with gelfoam sponges containing different proangiogenic factors and treated orally with water or PTK 787, a novel tyrosine kinase inhibitor with specific activity against the VEGF-R. PTK 787 significantly inhibited angiogenesis in sponges containing agarose + VEGF but not other proangiogenic molecules. The data show that the implanted gelfoam sponges provide a reliable quantitative assay to study in vivo angiogenesis.
McCarty
Marya F
MF
Department of Cancer Biology, The University of Texas M.D. Anderson Cancer Center, Houston, TX 77030, USA.
Baker
Cheryl H
CH
Bucana
Corazon D
CD
Fidler
Isaiah J
IJ
D020533:Angiogenesis Inhibitors / Q000494:pharmacology; D000818:Animals; D001681:Biological Assay / Q000379:methods*; D005260:Female; D005455:Fluorescent Antibody Technique; D005781:Gelatin Sponge, Absorbable; D006801:Humans; D066298:In Vitro Techniques; D051379:Mice; D008809:Mice, Inbred C3H; D009374:Neoplasms, Experimental / Q000098:blood supply* / Q000473:pathology; D018919:Neovascularization, Physiologic / Q000187:drug effects; D010793:Phthalazines / Q000494:pharmacology; D019408:Platelet Endothelial Cell Adhesion Molecule-1 / Q000378:metabolism; D011505:Protein-Tyrosine Kinases / Q000037:antagonists & inhibitors; D011725:Pyridines
D020533:Angiogenesis Inhibitors; D010793:Phthalazines; D019408:Platelet Endothelial Cell Adhesion Molecule-1; D011725:Pyridines; C404768:vatalanib; D011505:Protein-Tyrosine Kinases