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Spontaneous lipid vesicle fusion with electropermeabilized cells
Corinne Ramos1, David Bonato, Mathias Winterhalter
1Institut de Pharmacologie et de Biologie Structurale du CNRS, UMR 5089, 205, route de Narbonne, 31077 Cedex 4, Toulouse, France.
FEBS Letters
|May 9, 2002
Summary
Mammalian cell fusion with lipid vesicles is achieved using electropermeabilization and calcium. More metastable liposomes enhance fusion efficiency with electropulsated cells, suggesting a role in exocytosis.
Area of Science:
- Cell biology
- Biophysics
- Membrane science
Background:
- Cell fusion is a fundamental biological process.
- Lipid vesicles are widely used in drug delivery and cell studies.
- Electroporation is a method to permeabilize cell membranes.
Purpose of the Study:
- To investigate the fusion process between electropermeabilized mammalian cells and large unilamellar lipid vesicles.
- To determine the influence of liposome properties on fusion efficiency.
- To explore the role of membrane destabilization in cell-vesicle fusion.
Main Methods:
- Electropermeabilization of mammalian cells.
- Preparation of large unilamellar lipid vesicles with varying properties.
- Calcium-induced prepulse contact.
- Fluorescence-based monitoring of content mixing.
- Assessment of cell viability post-fusion.
Main Results:
- Fusion was successfully achieved and monitored using a fluorescence assay.
- Calcium ions promoted contact between cells and negatively charged liposomes.
- Fusion occurred under conditions that preserved cell viability.
- Liposome metastability correlated positively with fusion efficiency with electropulsated cells.
- Destabilization of only one membrane partner was sufficient for fusion.
Conclusions:
- Metastable liposomes exhibit higher fusability with electropulsated cells.
- Contact-induced electrostatic destabilization of the plasma membrane may be crucial for exocytosis.
- This study provides insights into membrane fusion mechanisms relevant to biological processes and biotechnology.