Loss of Thrombin-Induced Ca²⁺ Mobilization in a Subpopulation of Platelets during Storage

Summary

Thrombin-induced changes in cytosolic free Ca²⁺ ([Ca²⁺]_i) were studied in human platelets that had been stored for up to 6 days. Changes in [Ca²⁺]_i were measured with Indo-1-loaded platelets and quantitated with two different methods: (i) measurement of the changes in total fluorescence; (ii) measurement of the [Ca²⁺]_i changes in individual platelets in a flow cytometer, allowing the detection of non-responding platelets. The maximal concentration of [Ca²⁺]_i after stimulation with 0.5 U of thrombin/ml decreased from 544 ± 58 nM (mean ± SEM, n = 6) on day 0, to 276 ± 9 nM on day 3 and to 203 ± 23 nM on day 6. The percentage of platelets responding to 0.5 U of thrombin/ml declined from 90 ± 2% on day 0 to 72 ± 4% on day 3, and to 47 ± 8% on day 6. Nevertheless, also the responding platelets showed a decreased rise in [Ca²⁺]_i.

The study shows that during platelet storage a decrease in the rise in [Ca²⁺]_i upon thrombin stimulation occurs. This decrease is partly due to the formation of a subpopulation of platelets that is completely unresponsive and partly due to a decreased responsiveness in the remainder of the platelets; it is not due to a gradual decline in [Ca²⁺]_i rise in all platelets. This phenomenon provides new insight in the functional defect of stored platelets.

• References

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