Methods In Endothelial Cell Biology

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.

The participation of endothelial cells in various physiologic and pathologic processes has been hypothesized since before the turn of the century. However, until recently, direct evidence for endothelial involvement in these processes has been extremely difficult to obtain due to the inability to study endothelial cell function in vitro. Though the possibility of using cultured endothelial cells to study endothelial cell function in vitro was recognized many years ago, the inability to culture unambiguously identifiable endothelial cells limited investigators in their studies of endothelial function. As a result, the field of endothelial cell biology lay relatively fallow for many years. The development in the early 1970's of routine and easily implemented methods for culturing human endothelial cells and the demonstration that cultured endothelial cells synthesized a physiologically relevant protein, Factor VIII/von Willebrand Factor, quickly changed this state of affairs. Over the following decade the scope of endothelial cell research rapidly widened, spreading in a number of directions. First, methods were developed to culture endothelial cells from a variety of species. Second, methods were developed to culture endothelial cells from different organs and types of blood vessels (arteries, veins, and capillaries) within a single species. Third, and most important, investigators began using cultured endothelial cells as tools to study the potential involvement of endothelial cells in a wide assortment of biologically interesting processes. The net result has been a tremendous increase in our understanding of endothelial cell function.

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 aim of the Handbooks in Practical Animal Cell Biology is to provide practical workbooks for those involved in primary cell culture. Each volume addresses a different cell lineage, and contains an introductory section followed by individual chapters on the culture of specific differentiated cell types. The authors of each chapter are leading researchers in their fields and use their first-hand experience to present reliable techniques in a clear and thorough manner. Endothelial Cell Culture contains chapters on endothelial cells derived from 1) lung, 2) bone marrow, 3) brain, 4) mammary glands, 5) skin, 6) adipose tissue, 7) female reproductive system, and 8) synovium.

In the cardiovascular sciences, an increasing demand for the use of modern methods of cell biology has developed. The use of specific cell culture models of the various tissues involved is essential for most of these novel approaches. This book meets the demand for acomprehensive and easy accessible source for cell cul- ture methods in cardiovascular research as it was not previously available. The basic methods for cultures of cardiomyocytes (embryonic and adult), endothelial cells (micro- and macrovascular), smooth muscle cells and pericytes are described in detail by an international selection of experts. Special chapters discuss the use of growth factors and attachment substrates, techniques for co-cultures, cultures on permeable filter membranes and microcarrier cultures. The methodological descriptions are sufficiently detailed for an immediate application in the laboratory. All chapters also contain a critical evaluation of alternative approaches.

Endothelial dysfunction is broadly defined as a disruption of the balance between vasoactive mediators and a propensity towards an inflammatory state. This volume provides an overview of the fields of endothelial dysfunction and inflammation through the discussion of topics ranging from the molecular biology of activated endothelial cells to the endothelium in inflammatory disease and therapeutic approaches targeting endothelial dysfunction. Topics include: Heterogeneity of the endothelium during inflammation, oxidative stress and endothelial dysfunction, biology and regulation of nitric oxide in inflammatory pathologies, endothelial dysfunction in inflammatory diseases, such as diabetes and atherosclerosis and Clinical methods used to assess endothelial function. This book brings together basic and clinical research to assist the reader in bridging connections from bench-to-bedside. Written by expert researchers in the fields of endothelial biology, inflammation research and clinical science, it serves as a useful reference for academic and industrial researchers, clinicians, and trainees in the medical profession.

This detailed book builds on the foundation of lymphatic protocols with tools to aid in the interpretation of large datasets created by angiogenesis research. For example, this includes protocols to explore how transcriptional phenotyping can be achieved, both in terms of exploring tissue-specific heterogeneity to the use of database mining of existing datasets. Similarly, bioinformatic techniques run alongside protocols for transcript analysis, cell biology, pathology, as well as physiological and developmental angiogenesis models. 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, Angiogenesis: Methods and Protocols provides a meaningful compendium of practical approaches to angiogenesis and endothelial cell biology research.