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Routine Screening Method for Microparticles in Platelet Transfusions
Published on: January 31, 2018
Platelet microparticles are heterogeneous and highly dependent on the activation mechanism: studies using a new
Sílvia Perez-Pujol1, Paul H Marker, Nigel S Key
1Department of Medicine, University of Minnesota Medical School, Minneapolis, Minnesota, USA.
Background:
Platelet-derived microparticles (MPs) are believed to play an important role in coagulation and inflammatory disorders. Unfortunately, MP size renders them difficult to study and analyze by conventional flow cytometry.
Methods:
We analyzed and characterized platelet-derived MPs, using antibodies against the major surface glycoproteins (GP), the platelet activation antigen P-selectin (CD62P), and a marker of procoagulant activity (phosphatidylserine exposure). MPs were generated by exposure of platelets to thrombin receptor activating peptide (TRAP) or ionophore. Both agonists induced significant microvesiculation of platelets, and the resulting MPs were analyzed by a new digital flow cytometer: Becton-Dickinson FACSAria.
Results:
Membrane GPs were equally well represented in MPs generated by either reagent. In contrast, P-selectin was more intensely expressed in TRAP-MPs, while phosphatidylserine (PS) expression was markedly increased in ionophore-MPs. Two distinct populations of TRAP-MPs (one PS-positive and another PS-negative) were apparent. The latter characteristic facilitated sorting of MPs according to their PS exposure.
Conclusions:
The data presented herein show a significant improvement in the methodology applied until now to the characterization of MPs. The ability to characterize and sort MP subpopulations may help to resolve their contributions to normal and pathological functions.
Insights
This study introduces advanced digital flow cytometry for analyzing platelet-derived microparticles (MPs). New methods allow better characterization of MP subpopulations, aiding research into coagulation and inflammatory disorders.
Area of Science:
- Hematology
- Biotechnology
- Cell Biology
Background:
- Platelet-derived microparticles (MPs) are implicated in coagulation and inflammatory diseases.
- Conventional flow cytometry faces challenges in analyzing the small size of MPs.
Purpose of the Study:
- To characterize platelet-derived MPs using advanced digital flow cytometry.
- To improve methodologies for MP analysis and subpopulation sorting.
Main Methods:
- Platelet microparticles (MPs) were generated using thrombin receptor activating peptide (TRAP) or ionophore.
- MPs were analyzed using a digital flow cytometer (Becton-Dickinson FACSAria).
- Antibodies targeted surface glycoproteins, P-selectin (CD62P), and phosphatidylserine (PS) exposure.
Main Results:
- Both TRAP and ionophore induced significant microvesiculation.
- P-selectin expression was higher in TRAP-MPs; PS exposure was higher in ionophore-MPs.
- Two TRAP-MP populations (PS-positive and PS-negative) were identified, enabling sorting by PS exposure.
Conclusions:
- The study presents improved methodology for MP characterization.
- The ability to sort MP subpopulations can elucidate their roles in health and disease.
