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Isolation and Characterization of Microvesicles from Peripheral Blood
Published on: January 6, 2017
Isolation and Characterization of Microvesicles from Peripheral Blood
Kerstin Menck1, Annalen Bleckmann2, Matthias Schulz2
1Department of Hematology/Medical Oncology, University Medical Center Göttingen; kerstin.menck@inserm.fr.
Abstract:
The release of extracellular vesicles (EVs) including small endosomal-derived exosomes (Exos, diameter < 100 nm) and large plasma membrane-derived microvesicles (MVs, diameter > 100 nm) is a fundamental cellular process that occurs in all living cells. These vesicles transport proteins, lipids and nucleic acids specific for their cell of origin and in vitro studies have highlighted their importance as mediators of intercellular communication. EVs have been successfully isolated from various body fluids and especially EVs in blood have been identified as promising biomarkers for cancer or infectious diseases. In order to allow the study of MV subpopulations in blood, we present a protocol for the standardized isolation and characterization of MVs from peripheral blood samples. MVs are pelleted from EDTA-anticoagulated plasma samples by differential centrifugation and typically possess a diameter of 100 - 600 nm. Due to their larger size, they can easily be studied by flow cytometry, a technique that is routinely used in clinical diagnostics and available in most laboratories. Several examples for quality control assays of the isolated MVs will be given and markers that can be used for the discrimination of different MV subpopulations in blood will be presented.
Insights
This study presents a standardized protocol for isolating and characterizing microvesicles (MVs) from blood. This method enables the study of MVs as potential biomarkers for diseases.
Area of Science:
- Cell Biology
- Biochemistry
- Biotechnology
Background:
- Extracellular vesicles (EVs), including exosomes and microvesicles (MVs), are released by cells and mediate intercellular communication.
- EVs in blood are promising biomarkers for diseases like cancer and infections.
- Studying specific MV subpopulations in blood requires standardized isolation and characterization methods.
Purpose of the Study:
- To present a standardized protocol for the isolation and characterization of microvesicles (MVs) from human peripheral blood.
- To enable the study of MV subpopulations in blood for diagnostic applications.
Main Methods:
- Microvesicles (MVs) are isolated from EDTA-anticoagulated plasma using differential centrifugation.
- The protocol includes quality control assays for isolated MVs.
- Flow cytometry is utilized for characterization due to MVs' size (100-600 nm) and the technique's clinical availability.
Main Results:
- A reproducible protocol for isolating MVs from blood samples is established.
- Quality control assays are provided for validating MV isolation.
- Markers for discriminating different MV subpopulations are presented.
Conclusions:
- The developed protocol facilitates standardized isolation and characterization of blood MVs.
- This standardization is crucial for utilizing MVs as reliable biomarkers in clinical diagnostics.
- The protocol supports further research into MV-based disease detection and monitoring.
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