Study on Stress‐Mediated Behavior and Preparation of Giant Vesicles

Shimanouchi, Toshinori; Umakoshi, Hiroshi; Kuboi, Ryoichi

doi:10.1002/9780470511534.ch29

Cited by 2 publications

(3 citation statements)

References 19 publications

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“…We considered the expansion model in which SUVs, as nuclei, produced on the electrode, grow up to GVs (Figure b−d). Since the diameter change from SUVs as nuclei to GVs results in a decrease in membrane fluidity, a decrease in the k Gr value should occur during the expansion of SUVs to GVs according to Figure . This implies that the GV formation process involves some kinetic process, not consistent with the analysis as a single kinetic process like in Figure .…”

Section: Resultsmentioning

confidence: 99%

“…GVs were prepared according to a previous procedure. ,, The dried lipid was hydrated by the appropriate solution at 60 °C for 3 h. The floating lipid aggregates were sucked with a glass pipet several times. The lipid aggregates were dispersed, and we could observe GVs more than 1 μm in diameter by using photomicroscopy.…”

Section: Methodsmentioning

confidence: 99%

“…Giant vesicles (GVs) are cell-size liposomes and are known to be advantageous to the visualization of physiological phenomena. − Some preparation methods of GVs have been presented, such as electroformation, − the hydration method, − chemically induced vesiculation using a cell, , membrane perturbation effect by the protein molecule, and emulsification using a microchannel . However, its formation mechanism has not been clarified in details.…”

Section: Introductionmentioning

confidence: 99%

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Kinetic Study on Giant Vesicle Formation with Electroformation Method

2009

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Giant vesicles (GVs) composed of zwitterionic phospholipids were prepared by the electroformation method. The growth behavior of GVs was quantitatively analyzed as a first-order kinetics of the radius of GVs to obtain the apparent growth rate constant k(Gr). On the basis of the dependence of the k(Gr) value on the preparation temperature and the lipid composition of GVs, the membrane fluidity of vesicle membranes was found to be dominant in the growth behavior of GVs. The comparison of the k(Gr) values with the membrane fluidity for GVs suggested the validity of the model that the swelling of the lipid membrane could induce the growth of GVs.

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Section: Resultsmentioning

confidence: 99%

Section: Methodsmentioning

confidence: 99%

Section: Introductionmentioning

confidence: 99%

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Kinetic Study on Giant Vesicle Formation with Electroformation Method

2009

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Efficient Preparation of Giant Vesicles as Biomimetic Compartment Systems with High Entrapment Yields for Biomacromolecules

Kuroiwa

Fujita

Kobayashi

et al. 2012

Chemistry & Biodiversity

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The 'lipid-coated ice-droplet hydration method' was applied for the preparation of milliliter volumes of a suspension of giant phospholipid vesicles containing in the inner aqueous vesicle pool in high yield either calcein, α-chymotrypsin, fluorescently labeled bovine serum albumin or dextran (FITC-BSA and FITC-dextran; FITC=fluorescein isothiocyanate). The vesicles had an average diameter of ca. 7-11 μm and contained 20-50% of the desired molecules to be entrapped, the entrapment yield being dependent on the chemical structure of the entrapped molecules and on the details of the vesicle-formation procedure. The 'lipid-coated ice droplet hydration method' is a multistep process, based on i) the initial formation of a monodisperse water-in-oil emulsion by microchannel emulsification, followed by ii) emulsion droplet freezing, and iii) surfactant and oil removal, and replacement with bilayer-forming lipids and an aqueous solution. If one aims at applying the method for the entrapment of enzymes, retention of catalytic activity is important to consider. With α-chymotrypsin as first model enzyme to be used with the method, it was shown that high retention of enzymatic activity is possible, and that the entrapped enzyme molecules were able to catalyze the hydrolysis of a membrane-permeable substrate which was added to the vesicles after their formation. Furthermore, one of the critical steps of the method that leads to significant release of the molecules from the water droplets was investigated and optimized by using calcein as fluorescent probe.

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Study on Stress‐Mediated Behavior and Preparation of Giant Vesicles

Cited by 2 publications

References 19 publications

Kinetic Study on Giant Vesicle Formation with Electroformation Method

Kinetic Study on Giant Vesicle Formation with Electroformation Method

Efficient Preparation of Giant Vesicles as Biomimetic Compartment Systems with High Entrapment Yields for Biomacromolecules

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