Evidence map›Paper›PMID 31324885›Full record

ArticleScientific reports2019

Development of a quantitative method to measure EV uptake.

Víctor Toribio, Sara Morales, Soraya López-Martín, Beatriz Cardeñes, Carlos Cabañas, María Yáñez-Mó

Open access · goldAbstract read
In one paragraph

Article in Scientific reports, 2019. The graph could read no effect estimate from its abstract, so it casts no vote on the map. Cited by 40 papers.

0numbers the graph read from it
0cells of the map it votes in
40citing papers in PubMed
2.4field-weighted citation impact, top 10% of its field
1 · What the graph read from it

What it found

Each row is one number read from the abstract, on the scale the paper reported it, with its interval. Left of the dashed line favours the treatment, right favours the comparator. Under each row is the sentence it came from. New to these charts? A ten-minute tutorial.

The abstract states no effect estimate the extractor could read, or names no intervention and outcome on the map, so this paper lights no cell and moves no belief. It is still indexed, cited and linked below.

2 · The registry

The trial behind it

Trials whose registry record cites this paper, or whose number appears in the abstract. A trial that started after this paper was published is citing it as background, not reporting it.

Neither the registry nor the abstract names a trial number. If this is a trial report, that itself is worth knowing.

3 · Its place in the literature

Who cites it

40 citing papers in PubMed, 63 citations in OpenAlex.

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  17. The Exosome-like Vesicles ofInternational journal of molecular sciences · 2023
    Article
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  19. Metabolic reprogramming of hepatocytes byJHEP reports : innovation in hepatology · 2023
    Article
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4 · The record

Corrections and comments

PubMed lists nothing against this paper. Absence here is not a guarantee, only a check that was made.

5 · Who and what money

Authors and funding

6 authors at 3 institutions in 1 country.

Víctor ToribioDepartamento de Biología Molecular, Universidad Autónoma de Madrid (UAM), Madrid, Spain.
Sara MoralesInstituto de Investigación Sanitaria La Princesa (IIS-IP), Madrid, Spain.
Soraya López-MartínDepartamento de Biología Molecular, Universidad Autónoma de Madrid (UAM), Madrid, Spain.
Beatriz CardeñesCentro de Biología Molecular Severo Ochoa CSIC-UAM, Madrid, Spain.
Carlos CabañasCentro de Biología Molecular Severo Ochoa CSIC-UAM, Madrid, Spain.
María Yáñez-MóDepartamento de Biología Molecular, Universidad Autónoma de Madrid (UAM), Madrid, Spain. maria.yannez@uam.es.ORCID http://orcid.org/0000-0001-7484-2866
Centro de Biología Molecular Severo Ochoa · ESUniversidad Autónoma de Madrid · ESUniversidad Complutense de Madrid · ES

Funding

No grant is acknowledged in the PubMed record.

6 · The paper itself

Abstract

The outstanding potential of Extracellular Vesicles (EVs) in medicine, deserves a detailed study of the molecular aspects regulating their incorporation into target cells. However, because EV size lies below the limit of resolution of optical techniques, quantification together with discrimination between EV binding to the target cell and uptake is usually not completely achieved with current techniques. Human tetraspanins CD9 and CD63 were fused to a dual EGFP-Renilla-split tag. Subcellular localization and incorporation of these fusion proteins into EVs was assessed by western-blot and fluorescence microscopy. EV binding and uptake was measured using either a classical Renilla substrate or a cytopermeable one. Incubation of target cells expressing DSP2 with EVs containing the complementary DSP1 portion could not recover fluorescence or luciferase activity. However, using EVs carrying the fully reconstituted Dual-EGFP-Renilla protein and the cytopermeable Renilla luciferase substrate, we could distinguish EV binding from uptake. We provide proof of concept of the system by analysing the effect of different chemical inhibitors, demonstrating that this method is highly sensitive and quantitative, allowing a dynamic follow-up in a high-throughput scheme to unravel the molecular mechanisms of EV uptake in different biological systems.

Indexed as

Biological TransportBreast NeoplasmsCell Line, TumorExtracellular VesiclesFemaleGenes, ReporterGreen Fluorescent ProteinsHigh-Throughput Screening AssaysHumansImidazolesLuciferases, RenillaLuminescent MeasurementsNanoparticlesPyrazinesRecombinant Fusion ProteinsSensitivity and SpecificityCD63 protein, humanCD9 protein, humancoelenterazineenhanced green fluorescent proteinGreen Fluorescent ProteinsImidazolesLuciferases, RenillaPyrazinesRecombinant Fusion ProteinsTetraspanin 29Tetraspanin 30

Identifiers

PMID31324885
PMCPMC6642168
OpenAlexW2963659351

What OpenQuestion holds

Textmetadata
LicenceCC BY
Read underepoch 390

Registered trials

None linked

Read under generation 80e0d062 · epoch 390. Bibliography from PubMed, PubMed Central and OpenAlex; grants from NIH RePORTER; trial links from ClinicalTrials.gov; estimates, votes and beliefs from the OpenQuestion graph.