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Abstract
A flat perfusion chamber was developed to study the interaction of blood platelets
in flowing blood with cultured human vessel wall cells, their connective tissue matrix,
and isolated connective tissue components at defined shear rate conditions. A cover
slip covered with endothelial cells or extracellular matrix components was introduced
into the chamber. Laser-Doppler velocimetry showed a symmetrical flow profile at flow
rates between 50 and 150 ml/min (wall shear rate 300 to 1100 sec−1). Platelet deposition was estimated by using blood platelets labeled with indium-111
or by a morphometric method. Blood platelets did not adhere to endothelial cells at
wall shear rates of 765 sec−1 and the endothelial cells remained attached for at least 10 min of perfusion. In
preconfluent cultures of endothelial cells, blood platelets adhered to extracellular
material in areas between the cells. Removal of endothelial cells by treatment with
0.5% Triton X-100 induced increased platelet adherence with a preference for certain,
as yet unidentified, fibrillar structures of the extracellular matrix. Platelet adherence
to equine collagen was also studied after coating the cover slips by spraying of small
collagen droplets followed by air drying. Platelet adherence and the subsequent platelet
aggregate formation occurred predominantly along visible collagen fibers. These studies
showed that this perfusion chamber has a laminar and symmetrical flow allowing qualitative
and quantitative investigation of platelet interaction with endothelial cells, their
extracellular matrix, and pure connective tissue components. A variety of wall shear
rates and exposure times can be applied at controlled conditions without removing
cells or extracellular material.
Abbreviations:
phosphate-buffered saline ((PBS)), factor VIII-von Willebrand factor ((FVIII-VWF)), inner diameter ((Ø))To read this article in full you will need to make a payment
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Article info
Footnotes
☆The investigations were supported by grants 76.090 and 28.004 from the Foundation for Medical Research (FUNGO), which is subsidized by the Netherlands Organization for the Advancement of Pure Research (Z.W.O.) and by the Netherlands Heart Foundation.
Identification
Copyright
© 1983 Published by Elsevier Inc.
