Vesiculation of platelets during in vitro aging.
Membranous microparticles (MP) appearing in the supernatant plasma of stored platelet concentrates (PC) were analyzed by flow cytometry. Two populations of MP were arbitrarily delineated by light scatter as larger or smaller than 0.5 micron fluorescent beads. An estimate of MP concentration was obtained by adding a known amount of fluorescent beads to each sample before analysis of a set number of counts on the flow cytometer. The addition of platelet activation inhibitors (prostaglandin E-1, theophylline, and aprotinin) to the anticoagulant during preparation of PC combined with a reduction in surface area of the storage container caused approximately a 40% reduction in the number of MP appearing during storage relative to donor-matched controls. In addition, the inhibited concentrates had 84% less platelet factor 3 (PF3) activity in the supernatant and 61% less released lactic dehydrogenase. A reduction in surface area of the container in the controls partially offset these differences. A significant correlation was found (rs = .748) between PF3 levels and the concentration of larger MP. The inhibitors did not reduce the small number of MP found in stored platelet-poor plasma. Surface antigen analysis showed that the majority of MP in PC were platelet-derived; most were positive for glycoprotein (GP) IIbIIIa (73%) and/or for GPIb (43% to 46%). We conclude that procoagulant MP are released from platelets during storage as a result of platelet activation augmented by interaction of platelets with the bag wall.
Bode
A P
AP
Department of Clinical Pathology and Diagnostic Medicine, East Carolina University School of Medicine, Greenville, NC 27858.
Orton
S M
SM
Frye
M J
MJ
Udis
B J
BJ
D000527:Alprostadil / Q000494:pharmacology; D000954:Antigens, Surface / Q000032:analysis; D007611:Aprotinin / Q000494:pharmacology; D001792:Blood Platelets / Q000187:drug effects / Q000502:physiology* / Q000648:ultrastructure; D001793:Blood Preservation; D002232:Carbocyanines; D002462:Cell Membrane / Q000187:drug effects / Q000276:immunology / Q000648:ultrastructure*; D002470:Cell Survival; D005434:Flow Cytometry; D005456:Fluorescent Dyes; D006801:Humans; D007770:L-Lactate Dehydrogenase / Q000097:blood; D015539:Platelet Activation; D010977:Platelet Factor 3 / Q000378:metabolism; D010980:Platelet Membrane Glycoproteins / Q000032:analysis / Q000378:metabolism; D013806:Theophylline / Q000494:pharmacology; D013997:Time Factors
D000954:Antigens, Surface; D002232:Carbocyanines; D005456:Fluorescent Dyes; D010980:Platelet Membrane Glycoproteins; D010977:Platelet Factor 3; C007392:3,3'-dihexyl-2,2'-oxacarbocyanine; D007611:Aprotinin; D013806:Theophylline; D007770:L-Lactate Dehydrogenase; D000527:Alprostadil