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Updated: Jul 4, 2026
Techniques for the Analysis of Extracellular Vesicles Using Flow Cytometry
Published on: March 17, 2015
Flow cytometric analysis of cell membrane microparticles
Monique P Gelderman1, Jan Simak
1Laboratory of Cellular Hematology, CBER FDA, Rockville, MD, USA.
Abstract:
Cell membrane microparticles (MPs) are phospholipid microvesicles shed from the plasma membrane of most eukaryotic cells undergoing activation or apoptosis. The presence of MPs is common in healthy individuals. However, an increase in their release is a controlled event and is considered a hallmark of cellular alteration. Microparticles display cell surface proteins that indicate their cellular origin. In addition, they may also express other markers, e.g., markers of cellular activation. Elevated levels of circulating MPs are associated with various vascular pathologies and their pathogenic potential has been widely documented. MPs have been analyzed in plasma and cell cultures by means of flow cytometry or solid phase assays. Here we present a three-color flow cytometric assay for immunophenotyping of MPs in plasma. This assay has been used to study elevated counts of different phenotypes of circulating endothelial MPs in several hematological and vascular diseases. A modified version of this assay can also be used for MP analysis in blood products and cell cultures.
Insights
Cell membrane microparticles (MPs), released during cell activation or apoptosis, can indicate cellular changes. This study details a flow cytometry assay for analyzing MPs in plasma, aiding in diagnosing vascular diseases.
Area of Science:
- Cellular Biology
- Immunology
- Pathophysiology
Background:
- Cell membrane microparticles (MPs) are vesicles shed from cell membranes during activation or apoptosis.
- Increased MP levels signify cellular alterations and are linked to vascular pathologies.
- MPs express surface proteins indicating cellular origin and activation status.
Purpose of the Study:
- To present a three-color flow cytometric assay for immunophenotyping MPs in plasma.
- To enable the study of circulating endothelial MPs in various diseases.
Main Methods:
- Development of a three-color flow cytometry assay.
- Immunophenotyping of MPs in human plasma.
- Application of the assay to study endothelial MPs in hematological and vascular diseases.
Main Results:
- The assay allows for detailed immunophenotyping of MPs in plasma.
- Elevated counts of specific circulating endothelial MP phenotypes were observed in diseases.
- The assay is adaptable for analyzing MPs in blood products and cell cultures.
Conclusions:
- The developed flow cytometry assay is effective for analyzing circulating MPs.
- This method aids in understanding the role of MPs in vascular pathologies.
- The assay provides a valuable tool for research and clinical diagnostics involving MPs.
