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GENERAL ARTICLES

The Effects of Test Temperature and Storage Temperature on Platelet Aggregation: A Whole Blood In Vitro Study

Department of General Anesthesiology and Intensive Care (B), Vienna Medical University, Vienna, Austria

Address correspondence and reprint requests to Sibylle A. Kozek-Langenecker, MD, Department of General Anesthesiology and Intensive Care (B), Vienna Medical University Waehringer Guertel 18-20, 1090-Vienna, Austria. Address e-mail to sibylle.kozek{at}meduniwien.ac.at.

We systematically evaluated the effects of test temperature and storage temperature on platelet aggregation using flow cytometry and impedance aggregometry. Aliquots of citrated whole blood from 27 healthy adult male volunteers were stored at 37°C and 22°C. Aliquots were subjected to impedance aggregometry in response to collagen, adenosine diphosphate, ristocetin, and arachidonic acid performed at 22°C, 34°C, 37°C, and 40°C. The expression of activated fibrinogen receptor was determined on adenosine diphosphate-activated platelets at 22°C and 37°C by whole blood flow cytometry using PAC-1 for fluorescent staining. Aggregation induced by collagen, ristocetin, and arachidonic acid was not significantly different at the test temperatures of 34°C and 37°C but was significantly impaired at 22°C. In contrast, adenosine diphosphate-induced aggregation was significantly increased at both 34°C and 22°C. Hyperthermia exclusively impaired collagen-induced aggregation. Storage temperature of 22°C exclusively enhanced adenosine diphosphate- and collagen-induced aggregation compared with storage at 37°C. The binding of PAC-1 was enhanced at test temperatures below 37°C. Prewarming the antibody above 22°C significantly decreased binding. Our results suggest that mild hypothermic test conditions have no relevant effect, whereas profound hypothermia induces defects in adhesion, thromboxane generation, and activation. The pathomechanism for the increased response to adenosine diphosphate at mild and profound hypothermia remains unclear. Storage temperature considerably affects the aggregation response to the agonists adenosine diphosphate and collagen but not to arachidonic acid and ristocetin. Flow cytometry using the temperature-labile antibody PAC-1 fails to assess temperature effects on platelet aggregability.

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