Evaluation of Platelet Tests for Measurement of Cell Integrity^[*]

Summary

Various tests were evaluated for their capacity to differentiate between platelet suspensions with different degrees of cell damage. Those suspensions were prepared by simultaneous isolation of platelets from the same platelet-rich plasma (PRP) using the following procedures:

1. centrifugation at 4°C with EDTA

2. gel filtration in Tangen’s buffer

3. gel filtration in Ca²⁺-free Tyrode’s solution

4. gel filtration in Ca²⁺-free Tyrode followed by dehydration against polyethylene glycol 20,000 and

5. albumin density gradient centrifugation.

In these suspensions and in the original PRP the following parameters were studied: 1. morphology; 2. aggregability upon ADP addition; 3. platelet factor 3 availability; 4. uptake of ¹⁴C-serotonin and ³H-adenine; 5. metabolism of ³H-adenine and adenylate energy charge; 6. endogenous total ATP, ADP and serotonin and 7. lactate dehydrogenase (LDH) activity.

Quantitation of pseudopod formation in the light or electron microscope and log dose response studies for ADP-induced aggregation proved to be the most sensitive and reproducible of the tests studied. Additional information could be obtained from measurement of the ³H-label in the ATP and hypoxanthine-inosine fractions and calculation of the adenylate energy charge. Determination of platelet factor 3 availability or uptake studies of ¹⁴C-serotonin and ³H-adenine were less suitable for discriminating between cell suspensions. Data for total ATP and serotonin concentrations and LDH activity differed between the cell suspensions but instead of detecting various degrees of cell damage they reflected alterations in platelet population caused by the isolation procedures.

^* Reported in part at the meeting of the Working Party on Platelets of the International Committee on Thrombosis and Haemostasis (Cochairpersons: Drs. H. J. Day and M. B. Zucker), Paris, 1975.

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