Isolation and characterization of extracellular vesicle subpopulations from tissues

Crescitelli, Rossella; Lässer, Cecilia; Lötvall, Jan

doi:10.1038/s41596-020-00466-1

Cited by 293 publications

(288 citation statements)

References 93 publications

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“…We and others have previously shown that EVs can be isolated from different types of tissues, but these studies have mainly focused on determining the cargo and role of EVs from brain tissue [ 18 , 19 , 20 , 21 ] and tumor tissue [ 10 , 11 ]. The current study builds on these findings by showing for the first time that EVs can also be isolated from mouse lung tissue using our previously published protocol with some modifications [ 12 ]. Using this protocol, we could show that EVs released in the lung during allergen-induced airway inflammation are enriched in inflammation-associated proteins, which are related with the activation of immune cells and an excessive immune response.…”

Section: Discussionmentioning

confidence: 99%

“…EVs were isolated from the lung tissue according to our previously published protocol with some minor changes [ 12 ]. Briefly, the tissues were weighed, and RPMI-1640 medium (Sigma Aldrich) was added to each lung in the Petri dish, and the tissues were sliced into small pieces (approximately 2 mm × 2 mm).…”

Section: Methodsmentioning

confidence: 99%

“…( A ) Experimental model of OVA-induced allergic airway inflammation. ( B ) Handling and processing of the lung tissue according to our previous protocol [ 12 ]. ( C ) Protocol for the isolation of EVs from the lung tissue.…”

Section: Figurementioning

confidence: 99%

“…Notably, T2 (IL-4 and IL-13), and Th17 (IL-17A and TNF-α) cytokine-stimulated bronchial epithelial cells released EVs with significantly altered proteomes compared to non-activated cells. Additionally, we have recently published several studies using a protocol to isolate EVs directly from metastatic melanoma tissue samples [ 10 , 11 , 12 ].…”

Section: Introductionmentioning

confidence: 99%

“…Our hypothesis is that the phenotypes of lung EVs undergo significant changes during allergic airway inflammation. To test this, we used our previously published tissue-derived EV isolation protocol [ 12 ]. We harvested EVs from allergen-sensitized mice that were challenged with allergen or vehicle, and we performed quantitative proteome analysis to determine how the proteomes were altered in vivo during allergic asthma-like inflammation.…”

Section: Introductionmentioning

confidence: 99%

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Immune-Associated Proteins Are Enriched in Lung Tissue-Derived Extracellular Vesicles during Allergen-Induced Eosinophilic Airway Inflammation

Lässer

Kishino

Park

et al. 2021

IJMS

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Studying the proteomes of tissue-derived extracellular vesicles (EVs) can lead to the identification of biomarkers of disease and can provide a better understanding of cell-to-cell communication in both healthy and diseased tissue. The aim of this study was to apply our previously established tissue-derived EV isolation protocol to mouse lungs in order to determine the changes in the proteomes of lung tissue-derived EVs during allergen-induced eosinophilic airway inflammation. A mouse model for allergic airway inflammation was used by sensitizing the mice intraperitoneal with ovalbumin (OVA), and one week after the final sensitization, the mice were challenged intranasal with OVA or PBS. The animals were sacrificed 24 h after the final challenge, and their lungs were removed and sliced into smaller pieces that were incubated in culture media with DNase I and Collagenase D for 30 min at 37 °C. Vesicles were isolated from the medium by ultracentrifugation and bottom-loaded iodixanol density cushions, and the proteomes were determined using quantitative mass spectrometry. More EVs were present in the lungs of the OVA-challenged mice compared to the PBS-challenged control mice. In total, 4510 proteins were quantified in all samples. Among them, over 1000 proteins were significantly altered (fold change >2), with 614 proteins being increased and 425 proteins being decreased in the EVs from OVA-challenged mice compared to EVs from PBS-challenged animals. The associated cellular components and biological processes were analyzed for the altered EV proteins, and the proteins enriched during allergen-induced airway inflammation were mainly associated with gene ontology (GO) terms related to immune responses. In conclusion, EVs can be isolated from mouse lung tissue, and the EVs’ proteomes undergo changes in response to allergen-induced airway inflammation. This suggests that the composition of lung-derived EVs is altered in diseases associated with inflammation of the lung, which may have implications in type-2 driven eosinophilic asthma pathogenesis.

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Section: Discussionmentioning

confidence: 99%

Section: Methodsmentioning

confidence: 99%

Section: Figurementioning

confidence: 99%

Section: Introductionmentioning

confidence: 99%

Section: Introductionmentioning

confidence: 99%

See 3 more Smart Citations

Immune-Associated Proteins Are Enriched in Lung Tissue-Derived Extracellular Vesicles during Allergen-Induced Eosinophilic Airway Inflammation

Lässer

Kishino

Park

et al. 2021

IJMS

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Integrative analysis of circulating tumor cells (CTCs) and exosomes from small‐cell lung cancer (SCLC) patients: a comprehensive approach

Papakonstantinou,

Roumeliotou,

Pantazaka

et al. 2024

Molecular Oncology

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The increased metastatic ability of small‐cell lung cancer (SCLC) necessitates the identification of new prognostic biomarkers for clinical evaluation during the disease course. Our previous research highlighted the clinical relevance of transcription factor JunB (JUNB), C‐X‐C chemokine receptor type 4 (CXCR4), and programmed cell death 1 ligand 1 (PD‐L1) in breast and non‐small cell lung cancer (NSCLC) patients. In the current study, we examined these biomarkers in circulating tumor cells (CTCs) and plasma‐derived exosomes from 100 treatment‐naïve SCLC patients. CTCs were analyzed using the VyCAP system, whereas exosomes were characterized molecularly and transcriptomically. JUNB, CXCR4, and PD‐L1 were highly prevalent in CTCs. Patients exhibited significantly increased protein exosomal expression of JUNB and CXCR4 compared to healthy individuals. Overexpression of JUNB and CXCR4 in exosomes can distinguish patients from normal donors, offering an interesting tool for early diagnosis. The presence of JUNB and/or CXCR4 in CTCs correlated with significantly poorer overall survival. CXCR4 exosomal overexpression was associated with CTC presence and their phenotypes. Conclusively, a comprehensive analysis of CTCs and exosomes provides useful prognostic and potential diagnostic tools for SCLC patients.

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Current Technology for Production, Isolation, and Quality Control of Extracellular Vesicles

Han,

Ma,

et al. 2023

Biomedical Applications of Extracellular Vesicles

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Isolation and characterization of extracellular vesicle subpopulations from tissues

Cited by 293 publications

References 93 publications

Immune-Associated Proteins Are Enriched in Lung Tissue-Derived Extracellular Vesicles during Allergen-Induced Eosinophilic Airway Inflammation

Immune-Associated Proteins Are Enriched in Lung Tissue-Derived Extracellular Vesicles during Allergen-Induced Eosinophilic Airway Inflammation

Integrative analysis of circulating tumor cells (CTCs) and exosomes from small‐cell lung cancer (SCLC) patients: a comprehensive approach

Current Technology for Production, Isolation, and Quality Control of Extracellular Vesicles

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