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细胞功能测定

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Focal Cerebral Ischemia

Rodent models of focal cerebral ischemia have been extremely useful in elucidating pathomechanisms of human stroke. Most commonly, a monofilament is advanced through the internal carotid artery of rodents to occlude the origin of the middle cerebral artery thus leading to critical isc ...

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Extracorporeal Assays of Thrombosis

Platelet deposition, adhesion/aggregation, to the damaged vessel wall or atherosclerotic plaque components has shown to play a major role in hemostasis, thrombosis, and the development of atherosclerosis. Platelet-vessel wall interaction and thrombus formation is driven by b ...

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Platelet Life Span and Apoptosis

Like many nucleated mammalian cells, the life and death of the anucleate platelet is regulated by Bcl-2 family proteins. Platelets depend on Bcl-xL for survival. Bcl-xL maintains platelet viability by restraining the killer protein Bak. When Bak is unleashed, it triggers classical intrin ...

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A Live Cell Micro-imaging Technique to Examine Platelet Calcium Signaling Dynamics Under Blood Flow

The platelet is a specialized adhesive cell that plays a key role in thrombus formation under both physiological and pathological blood flow conditions. Platelet adhesion and activation are dynamic processes associated with rapid morphological and functional changes, with the ea ...

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Platelet Shape Change and Spreading

Hemostasis is dependent upon the successful recruitment and activation of blood platelets to the site of a breach in the vasculature. Platelet activation stimulates the rapid reorganization of the cortical actin cytoskeleton, resulting in the transformation of platelets from bic ...

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Clot Retraction

The study of clot retraction in vitro has been adopted as a simple and reproducible approach to assess platelet function. Plasma clots should retract away from the sides of a glass tube within a few hours allowing the rapid characterization of outside-in signaling through platelet integrin αI ...

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Visualization and Manipulation of the Platelet and Megakaryocyte Cytoskeleton

Driven by the application of immunofluorescence (IF) microscopy and modern molecular biology approaches to cytoskeletal manipulation, the last 5 years have yielded considerable progress to our understanding of the molecular mechanisms governing megakaryocyte developm ...

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Characterization of Megakaryocyte Development in the Native Bone Marrow Environment

Differentiation and maturation of megakaryocytes occur in close association with cellular and extracellular components in the bone marrow. Thus, direct examination of these processes in the native environment provides important information regarding the development of me ...

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Culture of Murine Megakaryocytes and Platelets from Fetal Liver and Bone Marrow

Megakaryocytes (MKs) are the largest hematopoietic cells in bone marrow. Since the mid-1990s, recombinant thrombopoietin has been commercially available. Together with the emerging knowledge of mouse models, this has provided an unprecedented contribution to the understand ...

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Platelet Proteomics: State of the Art and Future Perspective

Platelets pose unique challenges to cell biologists due to their lack of nucleus and low levels of messenger RNA. Platelets cannot be cultured in great abundance or manipulated using common recombinant DNA technologies. As a result, platelet research has lagged behind that of nucleated ce ...

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Endogenous Inhibitory Mechanisms and the Regulation of Platelet Function

The response of platelets to changes in the immediate environment is always a balance between activatory and inhibitory signals, the cumulative effect of which is either activation or quiescence. This is true of platelets in free flowing blood and of their regulation of haemostasis and thro ...

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Platelet Receptor Shedding

Receptor shedding is a mechanism for irreversible removal of transmembrane cell surface receptors by proteolysis of the receptor at a position near the extracellular surface of the plasma membrane. This process generates a soluble ectodomain fragment and a membrane-associated re ...

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Using Zebrafish (Danio rerio) to Assess Gene Function in Thrombus Formation

Cardiovascular and cerebrovascular disease is the major cause of death in the developed world, with a high burden of disease and substantial pharmaceutical investment to manage it (WHO, Global Burden of Disease, 2004 Update, W.H. Organisation, Editor. 2008). Platelets, as the principal me ...

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Megakaryopoiesis and Thrombopoiesis: An Update on Cytokines and Lineage Surface Markers

Megakaryopoiesis is the process by which mature megakaryocytes (MKs) develop from hematopoietic stem cells (HSCs). The biological function of MKs is to produce platelets, which play critical roles in hemostasis and contribute to angiogenesis and wound healing. The generation of pla ...

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Assessment of Megakaryocyte Migration and Chemotaxis

Cell migration is a highly integrated multistep process that plays an essential role during development and disease. Megakaryocytes (MKs) are specialized precursor cells that produce platelets and release them into the circulation. MK migration from the proliferative osteobl ...

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Purification of Native Bone Marrow Megakaryocytes for Studies of Gene Expression

Megakaryocytes constitute less than 1% of all marrow cells, therefore purification of these giant platelet precursor cells represents a challenge. We describe two methods to ultra-purify mature megakaryocytes from murine marrow for the purpose of extracting RNA suitable for studi ...

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Culture of Megakaryocytes and Platelets from Subcutaneous Adipose Tissue and a Preadipocyte Cell Line

The molecular mechanisms whereby stem cells develop into platelet-producing megakaryocytes (MKs) are not yet fully understood. Within this chapter we describe a two-step in vitro culture system in which MKs and platelets are generated from primary subcutaneous adipose tissues and t ...

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Megakaryocyte and Platelet Production from Human Cord Blood Stem Cells

The cloning of thrombopoietin together with advances in the culture of hematopoietic stem cells have paved the way for the study of megakaryopoiesis, ongoing clinical trials and, in the future, for the potential therapeutic use of ex vivo produced blood substitutes, such as platelets. This c ...

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In Vitro Generation of Megakaryocytes and Platelets from Human Embryonic Stem Cells and Induced Pluripotent Stem Cells

Human embryonic stem cells (hESCs) represent a potential source of blood cells for transfusion therapies and a promising tool for studying the ontogeny of hematopoiesis. Moreover, human-induced pluripotent stem cells (hiPSCs), recently established by defined reprogramming fa ...

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Mitochondrial DNA Mutations: An Overview of Clinical and Molecular Aspects

Mutations that arise in mitochondrial DNA (mtDNA) may be sporadic, maternally inherited, or Mendelian in character and include mtDNA rearrangements such as deletions, inversions or duplications, point mutations, or copy number depletion. Primary mtDNA mutations occur sporadi ...

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