Dynamic light scattering for the characterization and counting of extracellular vesicles: a powerful noninvasive tool

Palmieri, Valentina; Lucchetti, Donatella; Gatto, Ilaria; Maiorana, Alessandro; Marcantoni, Margherita; Maulucci, Giuseppe; Papi, Massimiliano; Pola, Roberto; Spirito, Marco De; Sgambato, Alessandro

doi:10.1007/s11051-014-2583-z

Cited by 108 publications

(95 citation statements)

References 24 publications

Supporting

Mentioning

Contrasting

Unclassified

Order By: Relevance

“…Size distribution of EV was measured by Dynamic light scattering (DLS) according to Palmieri et al using a Submicron Particle Sizer, Nicomp 380 instrument (Santa Barbara, CA), equipped with a solid‐state He‐Ne laser at 633 nm wavelength. The intensity of the scattered light was measured at 25°C.…”

Section: Methodsmentioning

confidence: 99%

Extracellular vesicles from oviductal isthmus and ampulla stimulate the induced acrosome reaction and signaling events associated with capacitation in bovine spermatozoa

Franchi

Moreno-Irusta

Domïnguez

et al. 2019

J of Cellular Biochemistry

View full text Add to dashboard Cite

Cells can communicate with other neighboring or distant cells through the secretion of extracellular vesicles (EV), composed of a lipid bilayer and bearing surface molecules that allow them to recognize target cells. In this way, EV induce signaling via different mechanisms, modulating the physiological state of the recipient cell. EV have been identified in both male and female reproductive fluids, however, the possible role of EV isolated from female reproductive fluids has become an emerging field only recently. It is known that ejaculated mammalian spermatozoa need to undergo physiological preparation in the female reproductive tract to fertilize the egg. EV secreted by different regions of the female tract constitute signals that may have a key role in regulating sperm functions. The aims of the present study were isolating EV from different regions of the bovine oviduct and analyzing their interaction and physiological effects on spermatozoa. Here, we report the characterization of bovine oviductal fluid EV from the isthmus and ampulla region and their effect on the induced acrosome reaction and signaling events associated with sperm capacitation. EV induced an increase in sperm protein tyrosine phosphorylation, while cell survival of cryopreserved bovine spermatozoa was maintained. We also show that EV uptake regulates the sperm calcium levels by inducing an immediate increase in the intracellular calcium concentration and sperm priming, after a pre‐incubation period, of the progesterone‐induced intracellular calcium rise. Our data contribute to understand the role of EV in the communication between the female reproductive tract and the sperm physiology, information that may be used to improve the efficiency of reproductive assisted technologies.

show abstract

Section: Methodsmentioning

confidence: 99%

Extracellular vesicles from oviductal isthmus and ampulla stimulate the induced acrosome reaction and signaling events associated with capacitation in bovine spermatozoa

Franchi

Moreno-Irusta

Domïnguez

et al. 2019

J of Cellular Biochemistry

View full text Add to dashboard Cite

show abstract

“…Indeed, US application can produce a series of biological effects at the cellular and tissue level that justifies its potential use in clinical practice (Xiong et al, 2015). In literature, numerous scientific evidence show that changes in the biomechanical and biophysical characteristics of cells are involved in the pathophysiology of many diseases, including cancer (Palmieri et al, 2014; Palmieri et al, 2015). For this reason, the study of the biological, structural, and molecular effects induced in cancer cells by the exposure to US is important to understand how and whether this tool might be suitable in the clinical practice.…”

Section: Introductionmentioning

confidence: 99%

Low‐intensity pulsed ultrasound affects growth, differentiation, migration, and epithelial‐to‐mesenchymal transition of colorectal cancer cells

Lucchetti

Perelli

Colella

et al. 2020

Journal Cellular Physiology

Self Cite

View full text Add to dashboard Cite

Ultrasound (US) offers potentially important opportunities from a therapeutic point of view. Thus, the study of the biological effects of US on cancer cells is important to understand the consequences of these changes on the malignant phenotype. This study aimed to investigate the effects of low‐intensity ultrasound (LIPUS) on the phenotype of colorectal cancer cell lines. Cell proliferation was evaluated by viability test and by evaluation of pERK expression, while cell motility using the scratch test. Cell differentiation was evaluated assessing alkaline phosphatase activity. Epithelial mesenchymal transition was assessed by analyzing the expression of Vimentin and E‐Cadherin. Release and uptake of extracellular vesicles (EVs) were evaluated by flow cytometry. LIPUS effects on the organization of cytoskeleton were analyzed by confocal microscopy and by evaluation of Rho GTPase expression. No alterations in vitality and clonogenicity were observed when the intermediate (0.4 MPa) and the lowest (0.035 MPa) acoustic intensities were administered while the treatment with high intensity (1 MPa) induced a reduction of both cell viability and clonogenicity in both cell lines in a frequency‐dependent manner. LIPUS promoted the differentiation of colon cancer cells, affected epithelial‐to‐mesenchymal transition, promoted the closure of a wound as well as increased the release of EVs compared with untreated cells. LIPUS‐induced increase in cell motility was likely due to a Rho GTPase‐dependent mechanism. Overall, the results obtained warrant further studies on the potential combined effect of LIPUS with differentiating agents and on their potential use in a clinical setting.

show abstract

“…One important issue is the determination of size distribution. Beside current techniques like electron microscopy (cryo-EM or freeze-fracture combined EM) [11,12], flow cytometry [13,14], and dynamic light scattering (DLS) [15], the introduction of novel techniques like nanoparticle tracking analysis (NTA) [16], resistive pulse sensing (RPS), small-angle X-ray scattering (SAXS) or size exclusion chromatography coupled with dynamic light scattering (SEC-DLS) are also ongoing [17,18]. Another key issue in EV research is the total protein determination and characterization by bioanalytical assays [19][20][21].…”

Section: Introductionmentioning

confidence: 99%

Characterization of extracellular vesicles by IR spectroscopy: Fast and simple classification based on amide and C H stretching vibrations

Mihály

Deák

Szigyártó

et al. 2017

Biochimica et Biophysica Acta (BBA) - Biomembranes

145

137

View full text Add to dashboard Cite

Extracellular vesicles isolated by differential centrifugation from Jurkat T-cell line were investigated by attenuated total reflection Fourier-transform infrared spectroscopy (ATR-FTIR). Amide and C-H stretching band intensity ratios calculated from IR bands, characteristic of protein and lipid components, proved to be distinctive for the different extracellular vesicle subpopulations. This proposed 'spectroscopic protein-to-lipid ratio', combined with the outlined spectrum-analysis protocol is valid also for low sample concentrations (0.15-0.05 mg/ml total protein content) and can carry information about the presence of other non-vesicular formations such as aggregated proteins, lipoproteins and immune complexes. Detailed analysis of IR data reveals compositional changes of extracellular vesicles subpopulations: second derivative spectra suggest changes in protein composition from parent cell towards exosomes favoring proteins with -turns and unordered motifs at the expense of intermolecular -sheet structures. The IR-based protein-to-lipid assessment protocol was tested also for red blood cell derived microvesicles for which similar values were obtained. The potential applicability of this technique for fast and efficient characterization of vesicular components is high as the investigated samples require no further preparations and all the different molecular species can be determined in the same sample. The results indicate that ATR-FTIR measurements provide a simple and reproducible method for the screening of extracellular vesicle preparations. It is hoped that this sophisticated technique will have further impact in extracellular vesicle research.

show abstract

Dynamic light scattering for the characterization and counting of extracellular vesicles: a powerful noninvasive tool

Cited by 108 publications

References 24 publications

Extracellular vesicles from oviductal isthmus and ampulla stimulate the induced acrosome reaction and signaling events associated with capacitation in bovine spermatozoa

Extracellular vesicles from oviductal isthmus and ampulla stimulate the induced acrosome reaction and signaling events associated with capacitation in bovine spermatozoa

Low‐intensity pulsed ultrasound affects growth, differentiation, migration, and epithelial‐to‐mesenchymal transition of colorectal cancer cells

Characterization of extracellular vesicles by IR spectroscopy: Fast and simple classification based on amide and C H stretching vibrations

Contact Info

Product

Resources

About