Culture-Produced Subendothelium

Booyse, François M.; Quarfoot, AJ; Feder, S

doi:10.1159/000214640

Cited by 8 publications

(1 citation statement)

References 9 publications

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“…The possibility of harvesting human endothelial cells (ECs) in the laboratory provides a useful tool for the investigation of the reactivity of vascular wall components towards platelets [1], The extracellular matrix (ECM) produced by ECs in culture is very similar to that found in basal membrane of the vascular wall [2], It is known that adhesive proteins such as von Willebrand factor (vWF), collagen, fibronectin or laminin [3][4][5][6], present in the ECM, mediate platelet interaction with the subendothelial wall [7], Exposure of ECM to circulat ing blood causes attachment of platelets [8] and in a short period of time circulating plate lets interact with previously attached platelets to form aggregates [9]. Initial attachment of platelets is mediated by vWF interacting with platelet GPIb [6] while the formation of aggre gates seems to be more dependent on fibrino gen interactions with platelet GPIIb-IIIa [ 10].…”

Section: Introductionmentioning

confidence: 99%

Digital Image Analysis of Platelet-Extracellular Matrix Interactions: Studies in von Willebrand Disease Patients and Aspirin-Treated Donors

Díaz‐Ricart¹,

Carretero²,

Castillo³

et al. 1994

Pathophysiol Haemos Thromb

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We have developed an automated image analysis method which provides en face morphometric evaluation of platelet interactions with coverslips coated with extracellular matrices. This procedure gives also additional information on the intensity of platelet-surface interactions in the same specimens. To standardize the method, anticoagulated human blood was flowed at 800 s^-1 during 10 min at 37°C through a parallel perfusion chamber in which extracellular matrix (ECM)-coated coverslips were placed. The sensitivity of the new method was tested in similar studies performed in patients with von Willebrand disease or treated with aspirin (500 mg/ day). The extent of platelet deposition on the ECMs was evaluated by both standard and new morphometric procedures. Coverslips were successively embedded in resin and processed for cross-sectional analysis and results were compared with those provided by the new automated procedure. A good correlation was found between results of covered surface obtained by both methods (r = 0.86; p < 0.005). Results from perfusions performed with blood from patients with platelet defects confirmed the correspondence of grey levels in the digitized images with the heights of groups of interacting platelets calculated from cross sections. The new automated method, in only one stage, allows a reliable evaluation of the global interaction of platelets with the ECM and effectively detected qualitative modifications in platelet-platelet interactions in patients with known platelet defects. The method described could be applied to the evaluation of the interactions of platelets with purified components of the subendothelium.

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Section: Introductionmentioning

confidence: 99%

Digital Image Analysis of Platelet-Extracellular Matrix Interactions: Studies in von Willebrand Disease Patients and Aspirin-Treated Donors

Díaz‐Ricart¹,

Carretero²,

Castillo³

et al. 1994

Pathophysiol Haemos Thromb

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Platelet-endothelial cell interactions in vitro following freeze-thaw injury or detergent treatment

1984

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Platelet interactions with cultured bovine endothelial cells were studied following freeze-thaw damage or detergent treatment. Platelets from whole blood, platelet-rich plasma, or gel-filtered plasma did not interact directly with freeze-thaw-damaged endothelial cells. Freezing and thawing did result in the exposure of an extracellular matrix located beneath the cells, which proved very thrombogenic. Platelets from all sources attached to both microfilament and amorphous components of the extracellular matrix, although only platelets from whole blood demonstrated aggregation and extensive pseudopodia formation. Treatment of cells with Triton-X detergent resulted in exposure of an intracellular cytoskeleton. Most platelets attached to the cytoskeleton were located near the cell border and had one or more pseudopodia either in contact with extracellular or intracellular material. Adhesion of platelets to the extracellular matrix may represent platelet-collagen or platelet-fibronectin interactions since both are produced by an incorporated into the extracellular matrix. Platelet interaction with endothelial cytoskeletons may represent contact of pseudopodia with the now exposed matrix located beneath the cells. The possibility that platelets also adhered to intracellular components could not be eliminated. These findings are in agreement with data from a freeze-thaw injury model of perfused aorta. In addition, they tend to indicate that physical insult is not sufficient to induce platelet interaction with the endothelial surface, but that chemical modification enhances platelet deposition.

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The relationship between von willebrand factor antigen and fibronectin in human plasma, endothelial cells and fibroblasts in culture

Giddings

Hogg

Legg

et al. 1986

Thrombosis Research

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Culture-Produced Subendothelium

Cited by 8 publications

References 9 publications

Digital Image Analysis of Platelet-Extracellular Matrix Interactions: Studies in von Willebrand Disease Patients and Aspirin-Treated Donors

Digital Image Analysis of Platelet-Extracellular Matrix Interactions: Studies in von Willebrand Disease Patients and Aspirin-Treated Donors

Platelet-endothelial cell interactions in vitro following freeze-thaw injury or detergent treatment

The relationship between von willebrand factor antigen and fibronectin in human plasma, endothelial cells and fibroblasts in culture

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