Novel Fusogenic Liposomes for Fluorescent Cell Labeling and Membrane Modification

Csiszár, Ágnes; Hersch, Nils; Dieluweit, Sabine; Biehl, Ralf; Merkel, Rudolf; Hoffmann, Bernd

doi:10.1021/bc900470y

Cited by 99 publications

(113 citation statements)

References 32 publications

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“…To assess the effect of lipids known to be enriched in caveolae, different Bodipy-labeled analogues of sphingolipids (Cer, SM C 5 and SM C 12 ), GSLs [ganglioside GM1 and lactosyl ceramide (LacCer)], Chol and phosphatidyl ethanolamine (PE) (Fig. S1B) were incorporated into liposomes [DOPE/Dotap/Bodipy-tagged lipid (47.5/47.5/5)] (Csiszar et al, 2010). Liposomes had diameters between 160-300 nm (Fig.…”

Section: Resultsmentioning

confidence: 99%

Lipid accumulation promotes scission of caveolae

Hubert

Larsson

Vegesna

et al. 2020

Preprint

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19Caveolae, bulb-shaped invaginations of the plasma membrane (PM), show distinct behaviors 20 of scission and fusion at the cell surface. Although it is known that caveolae are enriched in 21 cholesterol and sphingolipids, exactly how lipid composition influences caveolae surface 22 stability has not yet been elucidated. Accordingly, we inserted specific lipids into the PM of 23 cells via membrane fusion and studied acute effects on caveolae dynamics. We demonstrate 24 that cholesterol and glycosphingolipids specifically accumulate in caveolae, which decreases 25 their neck diameter and drives their scission from the cell surface. The lipid-induced scission 26 was counteracted by the ATPase EHD2. We propose that lipid accumulation in caveolae 27 generates an intrinsically unstable domain prone to scission if not balanced by the restraining 28 force of EHD2 at the neck. Our work advances the understanding of how lipids contribute to 29 caveolae dynamics, providing a mechanistic link between caveolae and their ability to sense 30 the PM lipid composition. 31 SUMMARY 32 Caveolae serve as mechanoprotectors and membrane buffers but their specific role in sensing 33 plasma membrane lipid composition remains unclear. Hubert et al. show that cholesterol and 34 glycosphingolipids accumulate in caveolae and drive subsequent scission from the cell 35 surface. These results provide new insight into how lipids contribute to budding and scission 36 of membrane domains in cells.37

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Section: Resultsmentioning

confidence: 99%

Lipid accumulation promotes scission of caveolae

Hubert

Larsson

Vegesna

et al. 2020

Preprint

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“…The CLVs possess unilamellar lipid bilayer membranes and can be produced with a wide range of uniform sizes through control of the flow-rates/applied pressures of the various component phases. After production, fragile molecules and biological components can be added to the surface for functionalization, or encapsulated into the SCMs using standard nano-vesicle fusion techniques 36,37 . Double emulsions can also be pre-functionalized or made to contain a stable drug compound before storage and then converted into CLVs at a later date when ready for use, allowing functional CLVs to be made in bulk and distributed to a wide variety of consumers for use as a cellular therapies, biosensors, and artificial cells.…”

Section: Resultsmentioning

confidence: 99%

Cell-sized lipid vesicles for cell-cell synaptic therapies

Vallejo

Lee

et al. 2017

Technology

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Cell-sized lipid vesicles (CLVs) have shown great promise for therapeutic and artificial cell applications, but their fragility and short shelf life has hindered widespread adoption and commercial viability. We present a method to circumvent the storage limitations of CLVs such as giant unilamellar vesicles (GUVs) and single-compartment multisomes (SCMs) by storing them in a double emulsion precursor form. The double emulsions can be stored for at least 8 months and readily converted into either GUVs or SCMs at any time. In this study, we investigate the interfacial parameters responsible for this morphological change, and we also demonstrate the therapeutic potential of CLVs by utilizing them to present a transmembrane protein, neuroligin-2, to pancreatic β-cells, forming cell-cell synapses that stimulate insulin secretion and cellular growth.

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“…We choose to work with the recently developed formulations reported by Csiszár et al because they were able to fuse with a large variety of cells. 29 Since the fusogenic activity of these systems is dependent on the use of uorescent lipids, fusion can be followed by the presence of uorescence in cells. Fig.…”

Section: Fusogenic Liposome Encapsulation Of Quantum Dotsmentioning

confidence: 99%

“…In this work, we adapted the freeze and thaw method to encapsulate water synthesized carboxyl-coated CdTe QDs in liposomes of different compositions. Recently, Csizár et al 29 reported direct fusion between non-functionalized cationic liposomes and the plasma membrane of different cell lineages. These liposomes with fusogenic properties are a valuable tool to investigate membrane fusion mechanisms and could also be used to design nanocarriers for intracellular delivery of membrane-impermeant compounds without the need for further chemical functionalization.…”

Section: Introductionmentioning

confidence: 99%

Studies on intracellular delivery of carboxyl-coated CdTe quantum dots mediated by fusogenic liposomes

et al. 2013

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The use of Quantum Dots (QDs) as fluorescent probes for understanding biological functions has emerged as an advantageous alternative over application of conventional fluorescent dyes. Intracellular delivery of QDs is currently a specific field of research. When QDs are tracking a specific target in live cells, they are mostly applied for extracellular membrane labeling. In order to study intracellular molecules and structures it is necessary to deliver free QDs into the cell cytosol. In this work, we adapted the freeze and thaw method to encapsulate water dispersed carboxyl-coated CdTe QDs into liposomes of different compositions, including cationic liposomes with fusogenic properties. We showed that labeled liposomes were able to fuse with live human stem cells and red blood cells in an endocytic-independent way. We followed the interactions of liposomes containing QDs with the cells. The results were minutely discussed and showed that QDs were delivered, but they were not freely diffused in the cytosol of those cells. We believe that this approach has the potential to be applied as a general route for encapsulation and delivery of any membrane-impermeant material into living cells.

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Novel Fusogenic Liposomes for Fluorescent Cell Labeling and Membrane Modification

Cited by 99 publications

References 32 publications

Lipid accumulation promotes scission of caveolae

Lipid accumulation promotes scission of caveolae

Cell-sized lipid vesicles for cell-cell synaptic therapies

Studies on intracellular delivery of carboxyl-coated CdTe quantum dots mediated by fusogenic liposomes

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