Over the past decades, the pathogenesis, diagnosis, treatment and prevention of cardiovascular diseases have been benefited significantly from intensive research activities. In order to provide a comprehensive “manual” in a field that has become as broad and deep as cardiovascular medicine, this volume of “Methods in Molecular Medicine” covers a wide spectrum of in vivo and in vitro techniques encompassing biochemical, pharmacological and molecular biology disciplines which are currently used to assess vascular disease progression. Each chapter included in this volume focuses on a specific vascular biology technique and describes various applications as well as caveats of these techniques. The protocols included here are described in detail, allowing beginners with little experience in the field of vascular biology to embark on new research projects.

Molecular biology has revolutionized research into vascular disease. Over the past 20 years molecular techniques have enabled us to both elucidate - lecular mechanisms in vascular disease and identify appropriate therapies. The vast explosion in technical knowledge and the array of protocols that become more advanced and intricate by the day lead us into new and exciting areas of research that were previously unobtainable. Vascular Disease: Molecular Biology and Gene Transfer Protocols - scribes today’s most powerful molecular methods for the investigation of the pathogenesis of vascular disease. The protocols are highly detailed, allowing beginners who have little experience in either vascular biology or molecular biology to embark on new molecular projects. This book is also suited to more experienced molecular biologists who wish to grasp new methods for stu- ing the involvement of genes in normal vascular physiology and in diseased states. It is well established that cardiovascular disease progression has a s- stantial genetic influence. Part I describes three methods that have been used successfully to identify specific mutations in candidate genes involved in c- diovascular disorders. These mutations include both single-stranded conf- mational polymorphism analysis and heteroduplex detection methods. In addition, technology to map new genes to specific regions of chromosomes by high-resolution mapping is described.

Over the past decades, the pathogenesis, diagnosis, treatment and prevention of cardiovascular diseases have been benefited significantly from intensive research activities. In order to provide a comprehensive “manual” in a field that has become as broad and deep as cardiovascular medicine, this volume of “Methods in Molecular Medicine” covers a wide spectrum of in vivo and in vitro techniques encompassing biochemical, pharmacological and molecular biology disciplines which are currently used to assess vascular disease progression. Each chapter included in this volume focuses on a specific vascular biology technique and describes various applications as well as caveats of these techniques. The protocols included here are described in detail, allowing beginners with little experience in the field of vascular biology to embark on new research projects.

A wide range of research methods for the study of vascular development, from basic laboratory protocols to advanced technologies used in clinical practice, are covered in this work. A range of methodologies such as molecular imaging platforms and signalling analysis, along with tumour models are collated here. Four sections explore in vitro techniques, in vivo and ex vivo manipulations, imaging and histological analysis and other novel techniques in vascular biology. Readers will discover basic methodologies used for analysis of endothelial cell growth in vitro, including co-culture models of vessel formation. Authors also explore isolation and purification of cells and methods for analysis of data and visualization of localized vasculature with modern imaging platforms. Both animal models and human disease are covered in this work. Each chapter contains helpful sections on trouble shooting, additional notes and links, supporting the reader to carry out protocols. This book will appeal to students, researchers and medical professionals working in all vascular-linked fields such as cardio- and cerebrovascular, cancer and dementia.

The lipid-rich and otherwise challenging nature of many key tissues complicates many aspects of current research, and applications of the unique nature of lipoproteins and their biological effects has engendered unique and vital methodologies. In Lipoproteins and Cardiovascular Disease: Methods and Protocols, experts in the field present a compendium of advanced and classical molecular biology methods targeted towards lipoprotein, atherosclerosis, and vascular biology research, bringing together in a single volume an updated set of protocols and strategies for methods now driving the most recent advances, along with classical methods that are still widely used. Among the many topics covered in this cutting-edge work, the book delves into crucial techniques such as quantitative real-time PCR, microarrays, RT-PCR laser capture microdissection, and tissue-specific gene overexpression, knockout, and knockdown methodologies, including AAV as a liver-directed gene delivery vehicle. Written in the highly successful Methods in Molecular BiologyTM series format, chapters include introductions to their respective subjects, lists of the necessary materials and reagents, step-by-step, readily reproducible laboratory protocols, and valuable notes which highlight tips on troubleshooting and avoiding known pitfalls. Comprehensive and easy to use, Lipoproteins and Cardiovascular Disease: Methods and Protocols serves both novices and experts alike as a complete guide for any researcher with an interest in lipoproteins and their significant biological effects.

This detailed new edition aims to provide a range of methods and protocols for studying vascular morphogenesis in vivo and in vitro to reflect up-to-date advances in the field. Of note, the book explores endothelial cell signaling is currently understood to promote fundamental cues for cell fate specification, embryo patterning, organ differentiation, and postnatal tissue remodeling. Written for the highly successful Methods in Molecular Biology series, chapters include introductions to their respective topics, lists of the necessary materials and reagents, step-by-step, readily reproducible laboratory protocols, and tips on troubleshooting and avoiding known pitfalls. Authoritative and timely, Vascular Morphogenesis: Methods and Protocols, Second Edition will serve as an ideal guide to this area of study for a broad audience amongst cell biologists, anatomists, pathologists, and physiologists.

This volume explores the latest techniques used to study the field of tissue engineered vascular grafts (TEVGs). The chapters in this book cover a wide array of topics such as deriving vascular cells from monocytes and induced pluripotent stem cells; engineering vascular grafts using various biomaterials and stem cells, stem cell-derived, or primary vascular cells; biomaterial modification by anticoagulation molecules; vascular bioengineering technologies such as 3D bioprinting; and fabrication of TEVGs with different geometry and multiphase structures. This book also features protocols for grafting and evaluation of vascular grafts in animal models, vascular imaging in animals, and the quantification of blood vessel permeability. Written in the highly successful Methods in Molecular Biology series format, chapters include introductions to their respective topics, lists of the necessary materials and reagents, step-by-step, readily reproducible laboratory protocols, and tips on troubleshooting and avoiding known pitfalls. Cutting-edge and practical, Vascular Tissue Engineering: Methods and Protocols is a valuable resource for biomedical engineers, cell biologists, vascular surgeons, doctors, and nurses.

This book explores techniques for exploring hydrogen sulfide (H2S) and its effects on the vascular system through numerous experimental animal models and vascular preparations. Alterations of vascular H2S generation/signaling may be involved in the pathogenesis of systemic and pulmonary arterial hypertension, ischemic heart disease, ischemic stroke, preeclampsia, and erectile dysfunction, and H2S also serves as an attractive target for pharmacotherapy of cardiovascular diseases, as well as possible effects on cancer, wound healing, and diabetic retinopathy, among other pathologies. Written for the highly successful Methods in Molecular Biology series, chapters include introductions to their respective topics, lists of the necessary materials and reagents, step-by-step, readily reproducible laboratory protocols, and tips on troubleshooting and avoiding known pitfalls. Authoritative and practical, Vascular Effects of Hydrogen Sulfide: Methods and Protocols is an ideal aid for scientists working to extend our knowledge in this valuable and wide-ranging field of study.

This volume provides an updated collection of protocols for manipulating and studying VEGF signaling pathways in vitro and in vivo and aims to present a range of both firmly established and newly emerging technologies. Covering multiple model species, from mouse to zebrafish to human, the book explores the role of VEGF and VEGFR isoforms in exosomes, cultured cells, or in tissues, as well as robust cell assays for the investigation of basic angiogenic mechanisms and VEGF signaling in more complex cellular systems, amongst other subjects. Written for the highly successful Methods in Molecular Biology series, chapters include introductions to their respective topics, lists of the necessary materials and reagents, step-by-step, readily reproducible laboratory protocols, and tips on troubleshooting and avoiding known pitfalls. Authoritative and up-to-date, VEGF Signaling: Methods and Protocols, Second Edition provides a useful tool for researchers in the vascular biology community and beyond in understanding the basic biology of VEGF signaling and in translating this research into the clinic.

In addition to updating important core techniques this third edition presents new chapters on assessing leukocyte involvement in angiogenesis, lymphatic cell and pericyte isolation techniques, spheroid and arterial ring based in vitro assays, and on pericyte involvement in angiogenesis. Angiogenesis Protocols, Third Edition also guides readers through new protocols on chorioallantoic membrane models, corneal pocket assays to assess angio- and lymphangiogenesis, models of muscle angiogenesis, and use of zebrafish embryos to study vascular angiogenesis and senescence. Written in the highly successful Methods in Molecular Biology series format, chapters include introductions to their respective topics, lists of the necessary materials and reagents, step-by-step, readily reproducible laboratory protocols, and tips on troubleshooting and avoiding known pitfalls. Authoritative and practical, Angiogenesis Protocols, Third Edition provides not only a practical handbook for key techniques, but also an informative and enjoyable read for all those interested, no matter how directly, in angiogenesis.