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EJ Harfenist, MA Guccione, MA Packham and JF Mustard
The method for preparing thrombin-degranulated platelets has been modified
to avoid the use of plasmin or successive treatments with small amounts of
thrombin, while still achieving more than 90% release of platelet amine
storage granule contents. It was necessary to prevent the fibrinogen
released from the platelets during thrombin treatment from forming an
insoluble fibrin mesh that could trap the platelets and hinder their
deaggregation. To accomplish this we have treated rabbit platelets with
0.73 U/ml of thrombin for 1 min in the presence of the synthetic peptide,
Gly-Pro-Arg-Pro, which prevents the polymerization of fibrin molecules. We
have demonstrated that it also prevents 125I, initially added as
125I-fibrinogen, from associating with the platelets in a form that was not
removed by centrifuging and washing during the preparation of
thrombin-degranulated platelets, and we infer that products formed from the
fibrinogen released from the platelets would also be prevented from
associating with them. Thrombin-degranulated platelets prepared by this
method have lost 92% of their granule contents and they can be washed and
resuspended. These platelets aggregate normally upon stimulation with
thrombin, adenosine diphosphate (ADP), or arachidonate. Thus,
Gly-Pro-Arg-Pro is useful in preparing thrombin-degranulated platelets for
studying platelet reactions without the complicating effects of released
materials such a ADP and fibrinogen.
This article has been cited by other articles:
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| Copyright © 1982 by American Society of Hematology Online ISSN: 1528-0020 | |||||||||
