Endothelial Cell Culture

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.

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.

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.

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.

This volume provides comprehensive protocols that researchers can use to dissect out fundamental principles and molecular mechanisms of hypertension, extending from genetics of experimental hypertension to biomarkers in clinical hypertension. Hypertension: Methods and Protocols guides readers through chapters on genetics and mmics of hypertension; the Renin-Angiotensin-Aldosterone System; vasoactive agents and hypertension; signal transduction and reactive oxygen species; novel cell models and approaches to study molecular mechanisms of hypertension; vascular physiology; and new approaches to manipulate mouse models to study molecular mechanisms of hypertension. 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 cutting-edge, Hypertension: Methods and Protocols aims to ensure successful results in the further study of this vital field.

Angiogenesis, the development of new blood vessels from the existing vasculature, is essential for physiological growth and over 18,000 research articles have been published describing the role of angiogenesis in over 70 different diseases, including cancer, diabetic retinopathy, rheumatoid arthritis and psoriasis. One of the most important technical challenges in such studies has been finding suitable methods for assessing the effects of regulators of eh angiogenic response. While increasing numbers of angiogenesis assays are being described both in vitro and in vivo, it is often still necessary to use a combination of assays to identify the cellular and molecular events in angiogenesis and the full range of effects of a given test protein. Although the endothelial cell - its migration, proliferation, differentiation and structural rearrangement - is central to the angiogenic process, it is not the only cell type involved. the supporting cells, the extracellular matrix and the circulating blood with its cellular and humoral components also contribute. In this book, experts in the use of a diverse range of assays outline key components of these and give a critical appraisal of their strengths and weaknesses. Examples include assays for the proliferation, migration and differentiation of endothelial cells in vitro, vessel outgrowth from organ cultures, assessment of endothelial and mural cell interactions, and such in vivo assays as the chick chorioallantoic membrane, zebrafish, corneal, chamber and tumour angiogenesis models. These are followed by a critical analysis of the biological end-points currently being used in clinical trials to assess the clinical efficacy of anti-angiogenic drugs, which leads into a discussion of the direction future studies should take. This valuable book is of interest to research scientists currently working on angiogenesis in both the academic community and in the biotechnology and pharmaceutical industries. Relevant disciplines include cell and molecular biology, oncology, cardiovascular research, biotechnology, pharmacology, pathology and physiology.