By Stewart Martin, Cliff Murray
A number of anti-angiogenic brokers at the moment are in medical trials for a number of ailments characterised by way of out of control blood vessel formation. This quantity offers a collection of up to date experiences at the vital elements of this method in addition to numerous options for learning angiogenesis.
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Extra resources for Angiogenesis Protocols
Screening Putative Angiogenic Compounds 1. Thaw Matrigel on ice overnight at 4°C. At no time should Matrigel be warmed to room temperature during handling. 2. Mix Matrigel on vortex; place on ice (see Notes 1 and 2). 3. Pipet equal amounts of Matrigel for each test condition in separate 14 mL tubes on ice. 5–1 mL with three or four mice for each condition (see Notes 2 and 3). 4. Tubes should be included for the positive and negative controls. Negative control is Matrigel alone; positive controls contain either 10 ng/mL and 100 ng/mL ECGS or 150 ng/mL bFGF added to Matrigel.
Schultz-Hector, S. and Haghayegh, S. (1993) Basic fibroblast growth factor expression in human and murine squamous cell carcinomas and its relationship to regional endothelial cell proliferation. Cancer Res. 53, 1444–1449. 69. Alvarez, J. , Gonzalez, A. , and Stopa, E. G. (1992) Localisation of basic fibroblast growth factor and vascular endothelial cell growth factor in human glial neoplasms. Mod. Pathol. 5, 303–307. 70. Guidi, A. , Abu, J. , Jackman, R. , Dvorak, H. , and Brown, L. F. (1995) Vascular permeability factor (vascular endothelial growth factor) expression and angiogenesis in cervical neoplasia.
5–1 mL, three or four mice for each condition (see Notes 2 and 3). 4. Add 150 ng/mL bFGF to all tubes except the one used for the negative control containing Matrigel alone. 5. Equalize the volumes with cold PBS so that, after test substances have been added, all tubes contain Matrigel at equal dilutions (see Note 4). Mix on the vortex. 6. 5 or 1 mL subcutaneously into the ventral area of each mouse (see Fig. 2; Notes 5 and 6). 7. Place animals of each test group in separate labeled cages. 8. After 7–10 d, sacrifice the animals and remove the plugs.