Improving extracellular vesicles visualization: From static to motion

Reclusa, Pablo; Verstraelen, Peter; Taverna, Simona; Gunasekaran, Muthukumar; Pucci, Marzia; Pintelon, Isabel; Claes, Nathalie; Pérez, Diego; Alessandro, Riccardo; Bals, Sara; Kaushal, Sunjay; Rolfo, Christian

doi:10.1038/s41598-020-62920-0

Cited by 35 publications

(25 citation statements)

References 51 publications

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“…For efficient HIGS, spray‐induced gene silencing (SIGS) can be employed to control eukaryotic microbial pathogens and plant‐feeding pests including nematodes and insects through application of pathogen or pest gene‐targeting sRNAs onto pre‐ and postharvest crops (Huang et al., 2019). Three‐dimensional EV tracking for in vivo internalization analysis of labelled EV uptake is anticipated to be realized in the field of plant pathology, as exemplified in animal cells (Liebel et al., 2020; Reclusa et al., 2020). Three‐dimensional analyses of the detailed processes of EV formation, trafficking and functionality will be addressed using high‐resolution imaging modalities, such as cryo‐electron tomography and cryo‐FIB.…”

Section: Discussionmentioning

confidence: 99%

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In situ and ex situ imaging of plant extracellular vesicles as nanovectors for cross‐domain communication

Kim

2021

Journal of Phytopathology

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Extracellular vesicles (EVs) are nanometric, lipid membrane‐bound vesicles released outside the cell. Plants have not been traditionally regarded to have EVs due to the presence of a cell wall. However, since the mid‐1960s, EVs have been reported in various plant species. They are often shown as vesicles in multivesicular bodies (MVBs) that later fuse with the plasma membrane (PM). Plant EVs have been commonly visualized using electron microscopy. Using membrane‐spanning domains, plant EVs can be fluorescence‐detected. They were observed near the infection sites in host tissues attacked by pathogens or at the interfaces with symbiotic fungi. Plant EVs could be taken up by bacteria and fungi. Proteins, lipids and nucleic acids, including small ribonucleic acids (RNAs), were found in plant EVs. Plant EVs have been isolated from the symplast and apoplast of various organs and purified after density gradient ultracentrifugation. Improved protocols and technologies for EV visualization provide a basis for the future applications of EVs in plant pathology.

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Section: Discussionmentioning

confidence: 99%

“…To minimize the time and procedure for animal EV purification, a commercial kit (exoEasy Maxi Kit; Qiagen) was used for SEM observations (Reclusa et al., 2020). The EVs isolated with the kit were fixed with 2.5% glutaraldehyde, dried on stubs and sputter‐coated with gold.…”

Section: Imaging Tools For Plant Evsmentioning

confidence: 99%

In situ and ex situ imaging of plant extracellular vesicles as nanovectors for cross‐domain communication

Kim

2021

Journal of Phytopathology

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“…Exosomes are known to play a very important role in the communication process between tumor cells and their microenvironment. Recently, several groups visualized through elegant imaging techniques the process of exosome uptake in NSCLC [31] and breast cancer [32]. The content of tumor-released exosomes can be uptaken by other adjoining tumor cells, tumor-niche (stroma) cells, immune cells, or distal organ cells after travelling through the circulatory system.…”

Section: Sending a Message: The Role Of Exosomes In Intercellular Communicationmentioning

confidence: 99%

Exosomes in Liquid Biopsy: The Nanometric World in the Pursuit of Precision Oncology

Valencia

Montuenga

2021

Cancers

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Among the different components that can be analyzed in liquid biopsy, the utility of exosomes is particularly promising because of their presence in all biological fluids and their potential for multicomponent analyses. Exosomes are extracellular vesicles with an average size of ~100 nm in diameter with an endosomal origin. All eukaryotic cells release exosomes as part of their active physiology. In an oncologic patient, up to 10% of all the circulating exosomes are estimated to be tumor-derived exosomes. Exosome content mirrors the features of its cell of origin in terms of DNA, RNA, lipids, metabolites, and cytosolic/cell-surface proteins. Due to their multifactorial content, exosomes constitute a unique tool to capture the complexity and enormous heterogeneity of cancer in a longitudinal manner. Due to molecular features such as high nucleic acid concentrations and elevated coverage of genomic driver gene sequences, exosomes will probably become the “gold standard” liquid biopsy analyte in the near future.

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“…2.7. SEM imaging of EVs of BMCM SEM imaging was performed to show EVs in the isolated BMCM (Reclusa et al, 2020). The isolated EVs were xed with 2.5% formalin buffer solution in PBS for 10 min.…”

Section: Isolation Of Evs Of Bmcm By Kitmentioning

confidence: 99%

Restoration of spermatogenesis in azoospermic mice by bone marrow mesenchymal stromal/stem cells conditioned medium

Zhankina

Afshar

Farrar

et al. 2021

Preprint

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One of the main cause of male infertility is non-obstructive azoospermia, which is not manageable medically. The first aim of the current research was to show the effect of extracellular vesicle-contained conditioned media (CM) instead of mesenchymal stromal/stem cells (MSCs) for treatment of non-obstructive azoospermia. In the next step, we aimed to study the differentiation potential of MSCs into spermatocytes after injection of MSCs in mice seminiferous tubules. This study has provided an applied treatment for busulfan-induced azoospermia using adipose tissue-derived (AT-MSCs) and bone marrow-derived MSCs (BM-MSCs) and bone marrow CM (BMCM) in animal models. In this regard, 30 male adult Balb/C mice (30±5g) and two female eGFP+/+ Balb/C mice (30±5g) were used to design experimental groups and to culture stem cells, respectively. Then, six groups including intact control, azoospermia, AT-MSC therapy, BM-MSC therapy, BMCM therapy, and spontaneous healing groups were considered. All groups except intact control were induced azoospermia by injecting two doses of busulfan (10 mg/kg) with 21 days’ interval. Testes of all mice were removed and studied through histomorphometry and flow cytometry analysis 60 days after treatment. Histomorphometry and flow cytometry evaluation of testes showed normal morphology of most of the seminiferous tubules of therapy groups as well as successful recovery of spermatogenesis, but spermatogenesis was not observed in the azoospermia group. It is worth notable that the results of the BM-MSC therapy group were more favorable than other therapy groups. Consequently, AT-MSC, BM-MSC and BMCM can be strongly suggested as candidates in the therapy of azoospermia.

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Improving extracellular vesicles visualization: From static to motion

Cited by 35 publications

References 51 publications

In situ and ex situ imaging of plant extracellular vesicles as nanovectors for cross‐domain communication

In situ and ex situ imaging of plant extracellular vesicles as nanovectors for cross‐domain communication

Exosomes in Liquid Biopsy: The Nanometric World in the Pursuit of Precision Oncology

Restoration of spermatogenesis in azoospermic mice by bone marrow mesenchymal stromal/stem cells conditioned medium

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