Evidence map›Paper›PMID 39044286›Full record

ArticleBMC research notes2024

Quality control and validation of extracellular vesicles isolated from cultured human breast cancer cells.

Urvi Patel, David Susman, Alison L Allan

Abstract readValidation Study
PubMed Full text at PMC Publisher
In one paragraph

Article in BMC research notes, 2024. The graph could read no effect estimate from its abstract, so it casts no vote on the map. Cited by 2 papers.

0numbers the graph read from it
0cells of the map it votes in
2citing papers in PubMed
–field-weighted citation impact
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

2 citing papers in PubMed.

  1. Review
  2. Review
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

3 authors.

Urvi PatelDepartment of Anatomy & Cell Biology, Western University, London, ON, N6A 3K7, Canada.
David SusmanDepartment of Anatomy & Cell Biology, Western University, London, ON, N6A 3K7, Canada.
Alison L AllanDepartment of Anatomy & Cell Biology, Western University, London, ON, N6A 3K7, Canada. alison.allan@lhsc.on.ca.

Funding

CIHR PJT-75100London Health Sciences Foundation N/A
6 · The paper itself

Abstract

objectiveExtracellular vesicles (EVs) have been shown to play a critical role in promoting tumorigenesis. As EV research grows, it is of importance to have standardization of isolation, quality control, characterization and validation methods across studies along with reliable references to explore troubleshooting solutions. Therefore, our objective with this Research Note was to isolate EVs from multiple breast cancer cell lines and to describe and perform protocols for validation as outlined by the list of minimal information for studies of EVs (MISEV) from the International Society for Extracellular Vesicles.

resultsTo isolate EVs, two techniques were employed: ultracentrifugation and size exclusion chromatography. Ultracentrifugation yielded better recovery of EVs in our hands and was therefore used for further validation. In order to satisfy the MISEV requirements, protein quantification, immunoblotting of positive (CD9, CD63, TSG101) and negative (TGFβ1, β-tubulin) markers, nanoflow cytometry and electron microscopy was performed. With these experiments, we demonstrate that yield of validated EVs varied between different breast cancer cell lines. Protocols were optimized to accommodate for low levels of EVs, and various technical and troubleshooting suggestions are included for potential application to other cell types that may provide benefit to investigators interested in future EV studies.

Indexed as

Breast NeoplasmsExtracellular VesiclesCell Line, TumorChromatography, GelDNA-Binding ProteinsEndosomal Sorting Complexes Required for TransportFemaleHumansQuality ControlTetraspanin 29Tetraspanin 30Transcription FactorsTsg101 ProteinUltracentrifugationDNA-Binding ProteinsEndosomal Sorting Complexes Required for TransportTetraspanin 29Tetraspanin 30Transcription FactorsTsg101 ProteinBreast cancerCell cultureElectron microscopyExtracellular vesiclesImmunoblottingNanoflow cytometryUltracentrifugation

Identifiers

PMID39044286
PMCPMC11265473

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Registered trials

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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.