| Contributors | p. xi |
| Preface | p. xv |
| Volumes in Series | p. xvii |
| Approaches for Studying Angiogenesis-Related Signal Transduction | p. 1 |
| Introduction | p. 2 |
| Angiogenesis | p. 2 |
| Phosphorylation of eNOS on Ser1179 | p. 8 |
| Measuring Released NO by No-Specific Chemiluminescence | p. 13 |
| Concluding Remarks | p. 18 |
| References | p. 19 |
| Physiologic Stress-Mediated Signaling in the Endothelium | p. 25 |
| Introduction | p. 26 |
| Parallel Plate Flow System | p. 29 |
| Cone and Plate Flow System | p. 35 |
| Cyclic Stretching of the Endothelium | p. 38 |
| Acknowledgments | p. 42 |
| References | p. 42 |
| Endothelial Cell Adhesion and Migration | p. 45 |
| Introduction | p. 46 |
| Cell Preparation | p. 48 |
| Preparation of Well-Defined Surfaces | p. 49 |
| Preparation of Polyacrylamide Substrates for Cell Adhesion Studies | p. 50 |
| Quantifying Cell Adhesion | p. 54 |
| Quantifying Endothelial Cell Migration | p. 56 |
| References | p. 61 |
| An Optimized Three-Dimensional In Vitro Model for the Analysis of Angiogenesis | p. 65 |
| Overview | p. 66 |
| Fibrin Bead Assay | p. 69 |
| Immunocytochemistry of Angiogenic Sprouts In Vitro | p. 73 |
| Retroviral Transduction of EC | p. 75 |
| Laser Capture Microdissection | p. 76 |
| Discussion | p. 78 |
| References | p. 79 |
| In Vitro Three Dimensional Collagen Matrix Models of Endothelial Lumen Formation During Vasculogenesis and Angiogenesis | p. 83 |
| Introduction | p. 84 |
| In Vitro Models of Vasculogenesis and Angiogenesis in 3D Collagen Matrices | p. 86 |
| Molecular/Genetic Manipulation of ECs | p. 93 |
| Rho GTPase Activation Assay During EC Tube Morphogenesis in 3D Collagen Matrices | p. 95 |
| Biochemical Analysis of Interactions Between Cdc42 and its Downstream Effectors During EC Lumen and Tube Formation in 3D Collagen Matrices | p. 96 |
| Conclusions | p. 98 |
| Acknowledgments | p. 99 |
| References | p. 99 |
| In Vitro Differentiation of Mouse Embryonic Stem Cells into Primitive Blood Vessels | p. 103 |
| Introduction | p. 104 |
| Maintenance of ES Cells | p. 107 |
| Generation of Transgenic ES Cell Lines | p. 111 |
| Concluding Remarks | p. 115 |
| Acknowledgments | p. 115 |
| References | p. 116 |
| The Aortic Ring Model of Angiogenesis | p. 119 |
| Introduction | p. 120 |
| Aortic Ring Angiogenesis Model: Basic Protocol | p. 122 |
| Protocols for Analysis of Aortic Cultures | p. 127 |
| Modifications of Basic Protocol | p. 129 |
| Troubleshooting | p. 132 |
| Concluding Remarks | p. 133 |
| References | p. 134 |
| An Assay System for In Vitro Detection of Permeability in Human "Endothelium" | p. 137 |
| Introduction | p. 138 |
| Generation of a Human Endothelium Model for Permeability Studies | p. 140 |
| Conclusion | p. 149 |
| Acknowledgments | p. 151 |
| References | p. 151 |
| Assays of Transendothelial Migration In Vitro | p. 155 |
| Introduction | p. 156 |
| Methods for Investigating Leukocyte Transendothelial Migration Under Static or Flow Conditions In Vitro | p. 156 |
| Acknowledgments | p. 174 |
| References | p. 174 |
| Cell Mechanics at Multiple Scales | p. 177 |
| Introduction | p. 178 |
| Large-Scale Cell Stretching | p. 179 |
| Magnetic Micromanipulation | p. 181 |
| Fluorescent Laser Tracking Microrheology | p. 186 |
| General Summary and Conclusion | p. 196 |
| Acknowledgments | p. 197 |
| References | p. 197 |
| Vascular Integrin Signaling | p. 199 |
| Introduction | p. 199 |
| Integrin-Mediated Cell Adhesion | p. 202 |
| Analysis of Integrin-Dependent Cell Migration | p. 204 |
| Roles of Integrins in Endothelial Angiogenic Programming | p. 210 |
| Integrin Affinity Modulation and Its Analysis | p. 217 |
| Integrin and ECM Roles in Endothelial Cell Proliferation and Apoptosis | p. 220 |
| Acknowledgment | p. 224 |
| References | p. 224 |
| Methods for Studying Mechanical Control of Angiogenesis by the Cytoskeleton and Extracellular Matrix | p. 227 |
| Introduction | p. 228 |
| Control of CE Cell Behavior with Different ECM Coating Densities | p. 229 |
| Control of CE Cell Fate Switching by Use of Microfabricated ECM Islands | p. 232 |
| Control of Cell Fate Switching by Modulating the Cytoskeleton | p. 240 |
| Analysis of the Mechanism of Cell Shape-Dependent Growth Control | p. 243 |
| Methods for Analyzing Rho-Dependent Control of Cell Growth and Movement | p. 246 |
| Rho-Dependent Control of Angiogenesis in Whole Organ Culture | p. 248 |
| Rho-Dependent Control of Vascular Permeability | p. 251 |
| Conclusion | p. 253 |
| Acknowledgments | p. 254 |
| References | p. 254 |
| VEGF Receptor Signal Transduction | p. 261 |
| Introduction | p. 262 |
| Methods | p. 264 |
| Conclusions | p. 282 |
| Acknowledgments | p. 283 |
| References | p. 283 |
| Analysis of Low Molecular Weight GTPase Activity in Endothelial Cell Cultures | p. 285 |
| Introduction | p. 285 |
| Methods and Results | p. 287 |
| Acknowledgments | p. 297 |
| References | p. 297 |
| Semaphorin-Induced Cytoskeletal Collapse and Repulsion of Endothelial Cells | p. 299 |
| Introduction | p. 300 |
| Purification of Human Recombinant SEMA3F | p. 301 |
| SEMA3F-induced Endothelial Cell Collapse | p. 305 |
| Endothelial Cell Repulsion Assay | p. 308 |
| References | p. 313 |
| Pericyte Isolation and Use in Endothelial/Pericyte Coculture Models | p. 315 |
| Introduction | p. 316 |
| Protocol for Isolating Pericytes | p. 319 |
| Protocol for Multipotent Mesenchymal Cells as Pericyte Precursors | p. 321 |
| Protocol for Two-Dimensional Pericyte-EC Coculture | p. 322 |
| Protocol for 3-Dimensional Pericyte-EC Coculture | p. 326 |
| Protocol for Differential Labeling of Pericytes and Endothelial Cells | p. 328 |
| Acknowledgments | p. 329 |
| References | p. 329 |
| Author Index | p. 333 |
| Subject Index | p. 351 |
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