[Platelet aggregation: a tool for clinical investigation and pharmacological study. Methodology]

Platelet aggregation can be measured quantitatively by continuous recording of the transmission of a beam of light across a suspension of platelets in constant agitation in an aggregometer. In routine clinical investigation, the study of platelet aggregation is performed on platelet-rich citrated plasma (PRPc). The blood sample has to be excellent to eliminate all traces of thrombin. The blood is collected in 3.8% sodium dihydrate citrate (1 volume for 9 volumes of blood). It is centrifuged at 190 g for 15 minutes at ambiant temperature. The PRPc obtained can be diluted with platelet-poor plasma (PPP) to adjust the concentration of platelets to 3 X 10(5)/microliters. The PRPc is stored at ambiant temperature in stoppered tubes under CO2 to avoid variations in pH. The maximal delay between the collection of the blood and the end of the study of aggregation is 3 hours. In certain cases, in order to define the platelet lesion and to eliminate the influence of plasma proteins, clotting factors and anti-coagulants, a suspension of washed platelets is used. The blood is collected on acid-citrate-dextrose (ACD) and centrifuged at + 37 degrees C to obtain the PRPc. The platelet deposit obtained by centrifugation of the PRPc is washed twice, according to Mustard's method, in Tyrode-albumin buffer at a concentration of 0.35% at + 37 degrees C. It is re-suspended in the same buffer solution at + 37 degrees C in the presence of apyrase and the platelet concentration is adjusted to 3 X 10(5)/microliters. The aggregation or agglutination of the platelets is induced by several agents: ADP, adrenalin, collagen, thrombin, arachidonic acid, ionophor A 23187, PAF-acether and ristocetin. The quantitative study of the aggregation curves of human platelets allows us to study the physiological and biochemical mechanisms control platelet aggregation, to recognize and classify the hereditary or acquired platelet abnormalities which lead to clinical haemorrhagic or thrombotic manifestations and to study the effect of drugs which inhibit platelet aggregation and to understand their mechanism of action.