Flow cytometric studies of platelet responses to shear stress in whole blood.

摘要: The objective of this work was to evaluate quantitatively the effects flow on platelet reactions using a cytometric technique. Whole blood exposed well defined, laminar shear stress in cone-and-plate viscometer absence added agonists. Blood specimens were fixed with formaldehyde and incubated two monoclonal antibodies. Antibody 6D1, specific for membrane glycoprotein Ib (GPIb), used identify enumerate platelets aggregates basis their characteristic forward scatter 6D1-FITC fluorescence profiles. Anti-CD62 antibody, granule protein-140 (GMP-140), measure activation. Results showed aggregation increasing marked increase response pathophysiological level 140 dyn/cm2 higher. This also apparent threshold formation large ("large" refers particles larger than 10 microns equivalent sphere diameter). These responses insensitive aspirin, but strongly inhibited by agents that elevate cyclic adenosine monophosphate (cAMP) levels. Moreover, pre-incubation whole antibodies inhibit von Willebrand factor binding GPIb or fibrinogen GPIIb/IIIa aggregation. Aggregation induced at 37 degrees C less extent 23 C. At physiological stresses, more susceptible shear-induced platelet-rich plasma. study reaffirms methods have several important advantages studies platelets, extends methodology unaltered cell separation methods.