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Updated: Aug 19, 2026
Isolation and Characterization of Microvesicles from Peripheral Blood
Published on: January 6, 2017
Plasma membrane vesiculation: a new technique for isolation of plasma membranes
Abstract:
Monolayer cell cultures of macrophages, monocytes, myoblasts, and density-inhibited and transformed fibroblasts form and release cell surface membrane vesicles following exposure to formaldehyde, related low-molecular-weight aldehydes, and disulfide blocking agents. Vesicles have a unique composition of proteins and lipids. They show enrichment of cholesterol and sphingomyelin content and a seven-to tenfold enrichment of 5'-nucleotidase activity. Vesicles also contain intramembranous particles and show a trilamellar unit membrane and no ultrastructural evidence of contamination with other cytoplasmic organelles. The technique is proposed as a novel method for isolating plasma membrane vesicles from cells in culture.
Insights
Researchers discovered a new method to isolate cell surface membrane vesicles from cultured cells using aldehydes and disulfide blockers. This technique yields unique vesicles, valuable for studying cell membranes.
Area of Science:
- Cell Biology
- Biochemistry
- Membrane Biology
Background:
- Cell surface membrane vesicles are crucial for intercellular communication and cellular processes.
- Existing methods for vesicle isolation can be complex and may lead to contamination.
- Understanding vesicle composition is key to elucidating their functions.
Purpose of the Study:
- To develop a novel method for isolating plasma membrane vesicles from various cultured cell types.
- To characterize the composition and ultrastructure of the isolated vesicles.
- To assess the utility of the technique for cell membrane research.
Main Methods:
- Exposure of cultured cells (macrophages, monocytes, myoblasts, fibroblasts) to formaldehyde, related aldehydes, and disulfide blocking agents.
- Isolation and biochemical analysis of released cell surface membrane vesicles.
- Ultrastructural examination of vesicle morphology and composition using electron microscopy.
Main Results:
- Aldehyde and disulfide blocking agent treatment induced the formation and release of cell surface membrane vesicles.
- Vesicles exhibited unique protein and lipid compositions, with enriched cholesterol and sphingomyelin.
- A significant enrichment of 5'-nucleotidase activity (7-10 fold) was observed in the vesicles.
- Ultrastructural analysis confirmed a trilamellar unit membrane structure and the presence of intramembranous particles, with no cytoplasmic organelle contamination.
Conclusions:
- The described method provides a novel and effective means for isolating plasma membrane vesicles from cultured cells.
- The unique composition of these vesicles makes them a valuable resource for studying membrane properties and functions.
- This technique offers a promising approach for advancing cell membrane research and understanding vesicle biology.
