Cell Biology


    Protocols in Current Issue
    Isolation and Characterization of Membrane Vesicles from Lactobacillus Species
    Authors:  Julie C. Caruana, Scott N. Dean and Scott A. Walper, date: 09/05/2021, view: 1772, Q&A: 0

    Throughout their life cycle, bacteria shed portions of their outermost membrane comprised of proteins, lipids, and a diversity of other biomolecules. These biological nanoparticles have been shown to have a range of highly diverse biological activities, including pathogenesis, community regulation, and cellular defense (among others). In recent

    Generation of Giant Unilamellar Vesicles (GUVs) Using Polyacrylamide Gels
    Authors:  Eric Parigoris, Daniel L. Dunkelmann and Unai Silvan, date: 11/05/2020, view: 2727, Q&A: 0
    [Abstract] Giant unilamellar vesicles (GUVs) are a widely used model system for a range of applications including membrane biophysics, drug delivery, and the study of actin dynamics. While several protocols have been developed for their generation in recent years, the use of these techniques involving charged lipid types and buffers of physiological ionic ...
    Buoyant Density Fractionation of Small Extracellular Vesicle Sub-populations Derived from Mammalian Cells
    Authors:  Morayma M. Temoche-Diaz, Matthew J. Shurtleff and Randy Schekman, date: 08/05/2020, view: 2969, Q&A: 0
    [Abstract] Small extracellular vesicles (sEVs) encompass a variety of distinct vesicles that are secreted to the extracellular space. Many methodologies currently used for EV isolation (e.g., differential ultracentrifugation concluding in a high-speed pellet, precipitation by macromolecular crowding agents or size excusion chromatography–SEC) do not ...
    Measuring Intracellular Vesicle Density and Dispersion Using Fluorescence Microscopy and ImageJ/FIJI
    [Abstract] Cell signalling, cell secretion, and plasma membrane repair are processes that critically rely on intracellular vesicles, important components of the endocytic and secretory pathways. More specifically, the strategic distribution of intracellular vesicles is important for diverse cellular processes. The method presented here is a simple, ...
    Isolating Multiple Extracellular Vesicles Subsets, Including Exosomes and Membrane Vesicles, from Bovine Milk Using Sodium Citrate and Differential Ultracentrifugation
    Authors:  Abderrahim Benmoussa, Sara Michel, Caroline Gilbert and Patrick Provost, date: 06/05/2020, view: 3673, Q&A: 0
    [Abstract] Milk is a complex fluid that contains various types of proteins and extracellular vesicles (EVs). Some proteins can mingle with EVs, and interfere with their isolation. Among these proteins, caseins form micelles of a size comparable to milk EVs, and can thus be co-isolated with EVs. Preliminary steps that affect milk are crucial for EV isolation ...
    Rapid Nickel-based Isolation of Extracellular Vesicles from Different Biological Fluids
    [Abstract] Extracellular vesicles (EVs) are membranous structures that cells massively release in extracellular fluids. EVs are cargo of cellular components such as lipids, proteins, and nucleic acids that can work as a formidable source in liquid biopsy studies searching for disease biomarkers. We recently demonstrated that nickel-based isolation (NBI) is a ...
    Study of Microbial Extracellular Vesicles: Separation by Density Gradients, Protection Assays and Labelling for Live Tracking
    [Abstract] Extracellular vesicles (EVs) are produced by all domains of life including Bacteria, Archaea and Eukarya. EVs are critical for cellular physiology and contain varied cargo: virulence factors, cell wall remodeling enzymes, extracellular matrix components and even nucleic acids and metabolites. While various protocols for isolating EVs have been ...
    Quantitative Analysis of Cargo Density in Single-extracellular Vesicles by Imaging
    Authors:  Taketoshi Kajimoto and Shun-ichi Nakamura, date: 12/20/2018, view: 4583, Q&A: 1
    [Abstract] Function of extracellular vesicles such as exosomes and microvesicles is determined by their wide ranges of cargoes inside them. Even in the pure exosomes or microvesicles the cargo contents are very heterogeneous. To understand this heterogeneous nature of extracellular vesicles, we need information of the vesicles, which will give us some ...
    Isolation and Quantification of Plant Extracellular Vesicles
    Authors:  Brian D. Rutter, Katie L. Rutter and Roger W. Innes, date: 09/05/2017, view: 17994, Q&A: 4
    [Abstract] Extracellular vesicles (EVs) play an important role in intercellular communication by transporting proteins and RNA. While plant cells secrete EVs, they have only recently been isolated and questions regarding their biogenesis, release, uptake and function remain unanswered. Here, we present a detailed protocol for isolating EVs from the ...

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