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Updated: Jan 31, 2026
Isolation and Characterization of Microvesicles from Peripheral Blood
Published on: January 6, 2017
Characterization of single microvesicles in plasma from glioblastoma patients
Kyle Fraser1, Ala Jo1, Jimmy Giedt1
1Center for Systems Biology, Massachusetts General Hospital Research Institute, Boston, Massachusetts.
Background:
Extracellular vesicles (EV) are shed by tumor cells but little is known about their individual molecular phenotypes and heterogeneity. While exosomes have received considerable attention, much less is known about larger microvesicles. Here we profile single microvesicles (MV) and exosomes from glioblastoma (GB) cells and MV from the plasma of patients.
Methods:
EV secreted from mouse glioma GL261 and human primary GBM8 cell lines as well as from the plasma of 8 patients with diagnoses of GB and 2 healthy controls were isolated and processed for single vesicle analysis. EV were immobilized on glass slides and the heterogeneity of vesicle and tumor markers were analyzed at the single vesicle level.
Results:
We show that (i) MV are abundant, (ii) only a minority of MV expresses putative MV markers, and (iii) MV share tetraspanin biomarkers previously thought to be diagnostic of exosomes. Using MV capture and staining techniques that allow differentiation of host cell and GB-derived MV we further demonstrate that (i) tumoral MV often present as <10% of all MV in GB patient plasma, and (ii) there is extensive heterogeneity in tumor marker expression in these tumor-derived MV.
Conclusion:
These results indicate that single MV analysis is likely necessary to identify rare tumoral MV populations and the single vesicle analytical technique used here can be applied to both MV and exosome fractions without the need for their separation from each other. These studies form the basis for using single EV analyses for cancer diagnostics.
Insights
Single microvesicle (MV) analysis reveals extensive heterogeneity in glioblastoma (GB) cell-derived extracellular vesicles (EV). This technique is crucial for identifying rare tumor-derived MV populations for cancer diagnostics.
Area of Science:
- Biochemistry
- Molecular Biology
- Oncology
Background:
- Extracellular vesicles (EVs) are shed by tumor cells, but their molecular phenotypes and heterogeneity remain poorly understood.
- While exosomes have been extensively studied, larger microvesicles (MVs) are less characterized.
- This study focuses on profiling single MVs and exosomes from glioblastoma (GB) cells and MVs from patient plasma.
Purpose of the Study:
- To investigate the molecular phenotypes and heterogeneity of single microvesicles (MVs) and exosomes from glioblastoma (GB) cells.
- To analyze MVs from the plasma of GB patients and healthy controls.
- To assess the utility of single EV analysis for cancer diagnostics.
Main Methods:
- EVs were isolated from mouse glioma GL261 and human primary GBM8 cell lines, as well as from the plasma of GB patients and healthy controls.
- Single vesicle analysis was performed by immobilizing EVs on glass slides.
- Heterogeneity of vesicle and tumor markers was analyzed at the single vesicle level using capture and staining techniques.
Main Results:
- Microvesicles (MVs) are abundant, but only a minority express putative MV markers.
- MVs share tetraspanin biomarkers previously considered specific to exosomes.
- Tumoral MVs constitute less than 10% of total MVs in GB patient plasma, exhibiting significant heterogeneity in tumor marker expression.
Conclusions:
- Single MV analysis is essential for identifying rare tumoral MV populations.
- The single vesicle analytical technique is applicable to both MV and exosome fractions without prior separation.
- These findings establish a foundation for utilizing single EV analyses in cancer diagnostics.
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