Methods in Endothelial Cell BiologyHellmut G. Augustin Endothelial cell biology has developed into a vibrant discipline and has become a critical instrument to study several disease processes on the cellular and molecular level. It is now widely recognized that dysfunctions of normal endothelial cell homeostasis are involved in some of the most important human diseases, including ischemic heart diseases, hypertension, atherosclerosis, tumors, diabetes, arthritis, and inflammation. Further, the increasing importance and recognition of the field of vascular biology in general requires in vitro and in vivo techniques in order to address the complex questions. Methods in Endothelial Cell Biology is a comprehensive practical "how-to"-guide summarizing the most relevant established techniques as well as a number of new emerging techniques. Easy-to-follow reliable protocols provide a useful lab bench resource for the experienced researcher and newcomer to the field. |
Contents
Isolation Purification and Culture of Human Micro and Macrovascular Endothelial Cells | 3 |
Isolation of Mouse Cardiac Microvascular Endothelial Cells | 15 |
Isolation Culture and Differentiation of Circulating Endothelial Precursor Cells | 23 |
Embryonic Stem CellDerived Endothelial Cells | 33 |
Detection Isolation and Culture of Lymphatic Endothelial Cells | 47 |
ThreeDimensional Spheroid Culture of Endothelial Cells | 55 |
Immortalization of Endothelial Cells | 63 |
Viral Gene Transfer Into Endothelial Cells | 73 |
Peripheral Hind Limb Ischemia Models | 197 |
In Vivo Matrigel Angiogenesis Assays | 207 |
Analysis of Vascular Permeability in Vivo | 223 |
Validating the Manipulation of Endothelial Cell Signaling in Vivo | 233 |
Chick Chorioallantoic Membrane Assay Growth Factor and Tumorinduced Angiogenesis and Lymphangiogenesis | 247 |
Corneal Angiogenesis Assay | 263 |
Detection of Endothelial Cells | 273 |
Basic Blood Vessel Immunohistochemistry | 275 |
Functional Assays in Vitro | 83 |
Leukocyte Adhesion to Endothelium in Vitro Under Shear Stress | 85 |
Application of Biomechanical Forces on Endothelial Cells | 93 |
Endothelial Cell Permeability Assays in Culture | 103 |
ThreeDimensional in Vitro Angiogenesis Assays | 115 |
Rat Aortic Ring Assay of Angiogenesis | 125 |
Endothelial Cell Chemotaxis and Chemokinesis Assays | 145 |
FACS Analysis of Endothelial Cells | 157 |
Application of the RNA Interference RNAi Technology to Angiogenesis Research | 167 |
Functional Assays in Vivo | 177 |
Murine Bone Marrow Transplantation Models that Enable the Study of EPC Recruitment | 179 |
Balloon Denudation of Blood Vessels | 187 |
DoubleStaining Immunohistochemistry of Blood Vessels | 287 |
In Situ Hybridization Analysis of Vascular Endothelium | 301 |
HighResolution in Situ Confocal Analysis of Endothelial Cells | 313 |
Whole Mount Analysis of the Embryonic Vasculature | 325 |
Methods for Visualizing Intact Blood Vessels Using Immunofluorescent Localization of PECAM CD31 and aSmooth Muscle Actin | 337 |
Intravital Multifluorescence Microscopy | 347 |
Corrosion Cast Analysis of Blood Vessels | 357 |
Appendix | 371 |
Suppliers | 373 |
Endothelial Cell Biology on the Web | 377 |
379 | |