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Applying flow cytometry-based isolation of platelet-derived microparticles to banked human plasma for cellular
Nima Fattahi1, Tarun Tyagi2, Fallyn Kirlin1
1Section of Digestive Diseases, Department of Internal Medicine, Yale University School of Medicine, New Haven, CT, USA.
Abstract:
Platelet-derived microparticles (PMPs) constitute the majority of circulating microparticles in blood. PMPs carry cargo including RNA, protein, and miRNA, and play key pathophysiologic intercellular signaling roles in cardiovascular, autoimmune, and liver diseases. Most methods use fresh platelets treated with an agonist to generate PMPs, which has restricted the size and feasibility of human studies. With the rise of large biobanks, such as the UK Biobank (https://www.ukbiobank.ac.uk/), and because microparticles are retained in cryopreserved plasma, this study aimed to develop an efficient and reproducible method to isolate PMPs from banked cryopreserved human plasma to investigate their communication with target cells. We employed flow cytometry using 180-1,300 nm size calibration beads and the platelet-specific marker CD41 to identify and sort PMPs based on size and CD41 positivity. Following isolation, microparticle size and morphology were validated by electron microscopy and nanoparticle tracking analysis. To assess the utility of the microparticles for studies of cell-cell interactions, we visualized microparticle uptake into human umbilical vein endothelial cells and THP-1 cells. Our method enables the isolation and downstream analysis of human PMPs in cell-cell interactions, facilitating translational studies in large populations and rare diseases. © 2026 The Pathological Society of Great Britain and Ireland.
Insights
Researchers developed a new method to isolate platelet-derived microparticles (PMPs) from cryopreserved plasma. This advance enables studying PMP communication in large biobanks and rare diseases.
Area of Science:
- Biomedical Science
- Cell Biology
- Hematology
Background:
- Platelet-derived microparticles (PMPs) are abundant in blood and mediate intercellular signaling in various diseases.
- Current PMP isolation methods using fresh platelets limit large-scale human studies.
- Cryopreserved plasma retains microparticles, offering a potential resource for research.
Purpose of the Study:
- To develop an efficient and reproducible method for isolating PMPs from banked cryopreserved human plasma.
- To enable downstream analysis of PMP communication with target cells.
- To facilitate translational research using large biobanks and studies of rare diseases.
Main Methods:
- Utilized flow cytometry with size calibration beads (180-1,300 nm) and CD41 marker for PMP identification and sorting.
- Validated PMP size and morphology using electron microscopy and nanoparticle tracking analysis.
- Assessed PMP uptake into human umbilical vein endothelial cells and THP-1 cells to evaluate functional utility.
Main Results:
- Successfully isolated and characterized PMPs from cryopreserved human plasma.
- Demonstrated the feasibility of analyzing PMP size, morphology, and cell uptake.
- Established a method compatible with large biobank samples.
Conclusions:
- The developed method efficiently isolates human PMPs from cryopreserved plasma.
- This technique supports the investigation of PMP-mediated cell-cell interactions.
- Enables translational research on PMPs in cardiovascular, autoimmune, and liver diseases.
