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You searched for: EV170041 (EV-TRACK ID)

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Experiment number
  • If needed, multiple experiments were identified in a single publication based on differing sample types, separation protocols and/or vesicle types of interest.
Species
  • Species of origin of the EVs.
Separation protocol
  • Gives a short, non-chronological overview of the different steps of the separation protocol.
    • (d)(U)C = (differential) (ultra)centrifugation
    • DG = density gradient
    • UF = ultrafiltration
    • SEC = size-exclusion chromatography
Details EV-TRACK ID Experiment nr. Species Sample type separation protocol First author Year EV-METRIC
EV170041 1/1 Homo sapiens Cell culture supernatant DG Yingkuan Shao 2018 87%

Study summary

Full title
All authors
Yingkuan Shao, Ting Chen, Xi Zheng, Sheng Yang, Kailun Xu, Xuewen Chen, Fei Xu, Lantian Wang, Yanwei Shen, Tingyang Wang, Mengwen Zhang, Wangxiong Hu, Chenyang Ye, XiaoFang Yu, Jimin Shao, Shu Zheng
Journal
Carcinogenesis
Abstract
Liver metastases develop in more than half of the patients with colorectal cancer (CRC) and are asso (show more...)Liver metastases develop in more than half of the patients with colorectal cancer (CRC) and are associated with a poor prognosis. The factors influencing liver metastasis of CRC are poorly characterized, but this information is urgently needed. We have now discovered that small extracellular vesicles (sEVs; exosomes) derived from CRC can be specifically targeted to liver tissue and induce liver macrophage polarization toward an interleukin-6 (IL-6)-secreting proinflammatory phenotype. More importantly, we found that microRNA-21-5p (miR-21) was highly enriched in CRC-derived sEVs and was essential for creating a liver proinflammatory phenotype and liver metastasis of CRC. Silencing either miR-21 in CRC-sEVs or Toll-like receptor 7 (TLR7) in macrophages, to which miR-21 binds, abolished CRC-sEVs' induction of proinflammatory macrophages. Furthermore, miR-21 expression in plasma-derived sEVs was positively correlated with liver metastasis in CRC patients. Collectively, our data demonstrate a pivotal role of CRC-sEVs in promoting liver metastasis by inducing an inflammatory premetastatic niche through the miR-21-TLR7-IL-6 axis. Thus, sEVs-miR-21 represents a potential prognostic marker and therapeutic target for CRC patients with liver metastasis. (hide)
EV-METRIC
87% (98th percentile of all experiments on the same sample type)
 Reported
 Not reported
 Not applicable
EV-enriched proteins
Protein analysis: analysis of three or more EV-enriched proteins
non EV-enriched protein
Protein analysis: assessment of a non-EV-enriched protein
qualitative and quantitative analysis
Particle analysis: implementation of both qualitative and quantitative methods
electron microscopy images
Particle analysis: inclusion of a widefield and close-up electron microscopy image
density gradient
Separation method: density gradient, at least as validation of results attributed to EVs
EV density
Separation method: reporting of obtained EV density
ultracentrifugation specifics
Separation method: reporting of g-forces, duration and rotor type of ultracentrifugation steps
antibody specifics
Protein analysis: antibody clone/reference number and dilution
lysate preparation
Protein analysis: lysis buffer composition
Study data
Sample type
Cell culture supernatant
Sample origin
Control condition
Focus vesicles
small extracellular vesicles
Separation protocol
Separation protocol
  • Gives a short, non-chronological overview of the
    different steps of the separation protocol.
    • (d)(U)C = (differential) (ultra)centrifugation
    • DG = density gradient
    • UF = ultrafiltration
    • SEC = size-exclusion chromatography
DG
Protein markers
EV: Alix/ HSP70/ CD63/ CD9
non-EV: Calreticulin
Proteomics
no
Show all info
Study aim
Function, Biomarker, Mechanism of uptake/transfer, Identification of content (omics approaches)
Sample
Species
Homo sapiens
Sample Type
Cell culture supernatant
Sample Condition
Control condition
EV-producing cells
'SW480 Dukes' type B, colorectal adenocarcinoma / SW620, colon / derived from metastatic site: lymph node
EV-harvesting Medium
EV-depleted serum
Preparation of EDS
overnight (16h) at >=100,000g
Cell viability
80
Separation Method
Density gradient
Only used for validation of main results
Yes
Density medium
Sucrose
Type
Discontinuous
Number of initial discontinuous layers
3
Lowest density fraction
0.25M
Highest density fraction
1.3M
Sample volume (mL)
1
Orientation
Bottom-up (sample migrates upwards)
Rotor type
SW 41 Ti
Speed (g)
100000
Duration (min)
150
Fraction volume (mL)
2
Fraction processing
Centrifugation
Pelleting: volume per fraction
5
Pelleting: duration (min)
150
Pelleting: rotor type
MLS-50
Pelleting: speed (g)
100000
Pelleting: adjusted k-factor
142.1
Pelleting-wash: volume per pellet (mL)
1
Pelleting-wash: duration (min)
150
Pelleting-wash: rotor type
69.83
Pelleting-wash: speed (g)
MLA-130
Pelleting-wash: adjusted k-factor
69.83
Characterization: Protein analysis
Protein Concentration Method
BCA
Protein Concentration
2-5
Western Blot
Antibody details provided?
Yes
Lysis buffer provided?
Yes
Detected EV-associated proteins
Alix, CD9, CD63, HSP70
Not detected contaminants
Calreticulin
Characterization: RNA analysis
Proteinase treatment
Moment of Proteinase treatment
After
Proteinase type
Proteinase K
Proteinase concentration
10
RNAse treatment
Moment of RNAse treatment
After
RNAse type
RNase A
RNAse concentration
10
Characterization: Particle analysis
NTA
Report type
Size range/distribution
Reported size (nm)
30-160
EV concentration
Yes
Particle yield
1.0-2.0E11
EM
EM-type
Transmission-EM
Image type
Wide-field
Report size (nm)
30-160
EV concentration
Yes
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