Optimization of Multiplate® whole blood platelet aggregometry in the Beagle dog and Wistar rat for ex vivo drug toxicity testing.

Abstract: This study was performed to optimize and standardize the use of the Multiplate^® whole blood impedance aggregometer in the Beagle dog and Wistar rat for use in a research laboratory environment. The anticoagulants citrate, heparin and hirudin were compared and platelet aggregation responses to ADP, collagen, arachidonic acid and Par-4 agonist were evaluated to determine their half maximal effective concentrations (EC₅₀) in blood containing low concentrations of a drug solvent (0.1% DMSO). The results indicate that citrate anticoagulation is not suitable for Multiplate^® whole blood aggregometry because of the presence of spontaneous aggregation. ADP and collagen were found to be appropriate agonists for both species, whereas in the Beagle dog Par-4 agonist failed to induce aggregation and arachidonic acid induced platelet aggregation showed a high interindividual variability. The agonists EC₅₀ calculated in hirudin blood were 2.70μM ADP, 0.85μg/ml collagen, 0.03mM arachidonic acid and 165.7μM Par-4 agonist in the Wistar rat, and 0.95μM ADP and 0.23μg/ml collagen in the Beagle dog. [Copyright &y& Elsevier]