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Related Experiment Videos

Platelet membrane integrity during storage and activation.

T J Reid1, G Esteban, M Clear

  • 1Department of Hematology and Vascular Biology, Walter Reed Army Institute of Research, Washington, DC 20307-5100, USA. reidt@wrsmtp-ccmail.army.mil

Transfusion
|June 23, 1999
PubMed
Summary

Dimethyl sulfoxide (DMSO) protects platelet membrane integrity during cooling and freezing, preventing cellular damage. This method using fluorescein diacetate (FD) offers a simple way to assess platelet membrane stability.

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Area of Science:

  • Biochemistry
  • Cell Biology
  • Hematology

Background:

  • Platelet cell membranes undergo lipid-phase transitions upon cooling, leading to cellular damage and material leakage.
  • Investigating protective agents for membranes against cooling injury is crucial for preserving platelet function.
  • Assessing cytosolic component leakage using fluorescein release is a key indicator of membrane integrity.

Purpose of the Study:

  • To evaluate the protective effects of various agents against cooling and freezing-induced platelet membrane damage.
  • To determine if agents beneficial for other cells can also protect platelets.
  • To establish a method for assessing platelet membrane integrity.

Main Methods:

  • Platelets were loaded with fluorescein diacetate (FD) and suspended in cryoprotective agents like DMSO or sugars.

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  • Platelets were subjected to cooling (4°C) or freezing (-70°C) and subsequent warming.
  • Fluorescein release, platelet aggregation, clot retraction, and p-selectin expression were measured to assess membrane integrity and function.

    • Main Results:

    • Dimethyl sulfoxide (DMSO) significantly reduced fluorescein release compared to other agents and chilling alone.
    • Platelet function remained largely unaffected by FD loading, indicating its suitability as a marker.
    • Fluorescein release correlated inversely with platelet function, confirming membrane damage.

    Conclusions:

    • Platelet loading with FD provides a straightforward method to evaluate membrane integrity.
    • DMSO demonstrates superior cryoprotective properties for platelets compared to other tested agents.
    • Maintaining platelet membrane integrity is essential for preserving their biological function.