Platelets and Megakaryocytes_ Volume 1_ Functional Assays - Ebook Pdf

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Platelets and Megakaryocytes_ Volume 1_ Functional Assays



Preface The average human body has in the order of 1012 circulating platelets. They are crucial for hemostasis, and yet excessive platelet activation is a major cause of morbidity and mortality in western societies. It is therefore not surprising that platelets have become one of the most extensively investigated biological cell types. We are, however, far from understanding precisely how platelets become activated under physiological and pathophysiological conditions. In addition, there are large gaps in our knowledge of platelet production from their giant precursor cell, the megakaryocyte. Understanding megakaryocyte biology will be crucial for the development of platelet gene targeting. The aim of Platelets and Megakaryocytes is therefore to bring together established and recently developed techniques to provide a comprehensive guide to the study of both the platelet and the megakaryocyte. It consists of five sections split between two volumes. The more functional assays appear in Volume 1, whereas Volume 2 includes signaling techniques, postgenomic methods, and a number of key perspectives chapters. Part I of Volume 1, Platelets and Megakaryocytes: Functional Assays, describes many well established approaches to the study of platelet function, including aggregometry, secretion, arachidonic acid metabolism, procoagulant responses, platelet adhesion under static or flow conditions, flow cytometry, and production of microparticles. Although one would ideally wish to perform experiments with human platelets, studies within the circulation using intravital microscopy require the use of animal models, which are described in Chapter 16, vol. 1. These approaches are becoming increasingly important in our understanding of how platelet responses contribute to the complex formation of thrombi within the circulation. Although naturally occurring genetic mutations can indicate the importance of specific proteins, these are limited in frequency and scope and thus many laboratories are using transgenic animals to delete or upregulate individual gene products (see Chapter 2, vol. 2). Consequently, the application of platelet techniques to murine models has become a focus of many labs in recent years (e.g., Chapters 2, 16, 20, vol. 1). In addition to basic and advanced approaches to study platelet function, several chapters in this section (particularly 1 and 2, vol. 1) focus on the long-standing issue of the effects of different anticoagulants and procedures to prepare platelets. The experimenter has a choice of studying platelets within the blood, in plasma, or in an artificial medium. In whole blood, potential interactions with other cell types and plasma proteins are included, which is in many ways the most physiological in vitro approach (see Chapter 6, vol. 1), however this is a complex situation and interpretation can be difficult. In studies within plasma, other cells are removed, but the clotting cascade is retained (see Chapter 5, vol. 1). Frequently, however, platelets are studied in isolation from other cells and plasma following their resuspension in an artificial medium. The preparation of platelets from human and other species is not a trivial matter and great care is required to ensure that the method of preparation does not adversely affect subsequent analysis (see Chapter 2, vol. 1).

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