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You searched for: EV170016 (EV-TRACK ID)

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Experiment number
  • If needed, multiple experiments were identified in a single publication based on differing sample types, isolation protocols and/or vesicle types of interest.
Species
  • Species of origin of the EVs.
Isolation protocol
  • Gives a short, non-chronological overview of the different steps of the isolation protocol.
    • dUC = differential ultracentrifugation
    • DG = density gradient
    • UF = ultrafiltration
    • SEC = size-exclusion chromatography
Experiment number
  • Experiments differ in Sample type
Experiment number
  • Experiments differ in Sample type
Details EV-TRACK ID Experiment nr. Species Sample type Isolation protocol First author Year EV-METRIC
EV170016 1/2 Mus musculus Cell culture supernatant DG
Filtration
dUC
Ying, Wei 2017 88%

Study summary

Full title
All authors
Ying W, Riopel M, Bandyopadhyay G, Dong Y, Birmingham A, Seo JB, Ofrecio JM, Wollam J, Hernandez-Carretero A, Fu W, Li P, Olefsky JM
Journal
Cell
Abstract
MiRNAs are regulatory molecules that can be packaged into exosomes and secreted from cells. Here, we (show more...)MiRNAs are regulatory molecules that can be packaged into exosomes and secreted from cells. Here, we show that adipose tissue macrophages (ATMs) in obese mice secrete miRNA-containing exosomes (Exos), which cause glucose intolerance and insulin resistance when administered to lean mice. Conversely, ATM Exos obtained from lean mice improve glucose tolerance and insulin sensitivity when administered to obese recipients. miR-155 is one of the miRNAs overexpressed in obese ATM Exos, and earlier studies have shown that PPARγ is a miR-155 target. Our results show that miR-155KO animals are insulin sensitive and glucose tolerant compared to controls. Furthermore, transplantation of WT bone marrow into miR-155KO mice mitigated this phenotype. Taken together, these studies show that ATMs secrete exosomes containing miRNA cargo. These miRNAs can be transferred to insulin target cell types through mechanisms of paracrine or endocrine regulation with robust effects on cellular insulin action, in vivo insulin sensitivity, and overall glucose homeostasis. (hide)
EV-METRIC
88% (99th 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
Isolation method: density gradient, at least as validation of results attributed to EVs
EV density
Isolation method: reporting of obtained EV density
ultracentrifugation specifics
Isolation 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
Focus vesicles
exosome
Isolation protocol
Isolation protocol
  • Gives a short, non-chronological overview of the
    different steps of the isolation protocol.
    • dUC = differential ultracentrifugation
    • DG = density gradient
    • UF = ultrafiltration
    • SEC = size-exclusion chromatography
DG + Filtration + dUC
Adj. k-factor
256 (pelleting) / 256 (washing)
Protein markers
EV: CD9/ CD63/ HSP70/ TSG101/ Syntenin1
non-EV: Grp94
Proteomics
no
Show all info
Study aim
Function, Mechanism of uptake/transfer
Sample
Species
Mus musculus
Sample Type
Cell culture supernatant
Sample Condition
Obesitas
EV-producing cells
adipose tissue-derived macrophages
EV-harvesting Medium
EV-depleted serum
Preparation of EDS
Commercial EDS
Isolation Method
Differential ultracentrifugation
dUC: centrifugation steps
Between 800 g and 10,000 g
Between 100,000 g and 150,000 g
Pelleting: time(min)
240-360
Pelleting: rotor type
SW 32 Ti
Pelleting: speed (g)
100000
Pelleting: adjusted k-factor
256.0
Wash: time (min)
20
Wash: Rotor Type
SW 32 Ti
Wash: speed (g)
100000
Wash: adjusted k-factor
256.0
Density gradient
Only used for validation of main results
Yes
Density medium
Iodixanol
Type
Discontinuous
Lowest density fraction
0.1
Highest density fraction
0.3
Sample volume (mL)
0.3
Orientation
Bottom-up (sample migrates upwards)
Rotor type
Type 70 Ti
Speed (g)
350000
Duration (min)
60
Fraction volume (mL)
2.4
Fraction processing
Centrifugation
Pelleting: volume per fraction
2
Pelleting: duration (min)
90
Pelleting: rotor type
Type 70 Ti
Pelleting: speed (g)
100000
Pelleting: adjusted k-factor
156.9
Pelleting-wash: volume per pellet (mL)
24
Pelleting-wash: duration (min)
30
Pelleting-wash: rotor type
156.9
Pelleting-wash: speed (g)
Type 70 Ti
Pelleting-wash: adjusted k-factor
156.9
Filtration steps
0.22µm or 0.2µm
EV-subtype
Distinction between multiple subtypes
1.13-1.15 g/ml
Used subtypes
Yes
Characterization: Protein analysis
Protein Concentration Method
DC protein assay
Protein Concentration
9-May
Western Blot
Antibody details provided?
Yes
Lysis buffer provided?
Yes
Detected EV-associated proteins
CD9, CD63, HSP70, TSG101, Syntenin1
Not detected contaminants
Grp94
Fluorescent NTA
Characterization: Particle analysis
NTA
Report type
Size range/distribution
Reported size (nm)
30-150
EM
EM-type
Transmission-EM
Image type
Wide-field
Report size (nm)
50-200 nm
Extra information
Full UC and density gradient protocols not in original article
EV170016 2/2 Mus musculus Cell culture supernatant dUC
Filtration
Ying, Wei 2017 55%

Study summary

Full title
All authors
Ying W, Riopel M, Bandyopadhyay G, Dong Y, Birmingham A, Seo JB, Ofrecio JM, Wollam J, Hernandez-Carretero A, Fu W, Li P, Olefsky JM
Journal
Cell
Abstract
MiRNAs are regulatory molecules that can be packaged into exosomes and secreted from cells. Here, we (show more...)MiRNAs are regulatory molecules that can be packaged into exosomes and secreted from cells. Here, we show that adipose tissue macrophages (ATMs) in obese mice secrete miRNA-containing exosomes (Exos), which cause glucose intolerance and insulin resistance when administered to lean mice. Conversely, ATM Exos obtained from lean mice improve glucose tolerance and insulin sensitivity when administered to obese recipients. miR-155 is one of the miRNAs overexpressed in obese ATM Exos, and earlier studies have shown that PPARγ is a miR-155 target. Our results show that miR-155KO animals are insulin sensitive and glucose tolerant compared to controls. Furthermore, transplantation of WT bone marrow into miR-155KO mice mitigated this phenotype. Taken together, these studies show that ATMs secrete exosomes containing miRNA cargo. These miRNAs can be transferred to insulin target cell types through mechanisms of paracrine or endocrine regulation with robust effects on cellular insulin action, in vivo insulin sensitivity, and overall glucose homeostasis. (hide)
EV-METRIC
55% (91st 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
Isolation method: density gradient, at least as validation of results attributed to EVs
EV density
Isolation method: reporting of obtained EV density
ultracentrifugation specifics
Isolation 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
Focus vesicles
exosome
Isolation protocol
Isolation protocol
  • Gives a short, non-chronological overview of the
    different steps of the isolation protocol.
    • dUC = differential ultracentrifugation
    • DG = density gradient
    • UF = ultrafiltration
    • SEC = size-exclusion chromatography
dUC + Filtration
Adj. k-factor
256 (pelleting) / 256 (washing)
Protein markers
EV: CD9/ CD63/ HSP70/ TSG101/ Syntenin1
non-EV: Grp94
Proteomics
no
Show all info
Study aim
Function, Mechanism of uptake/transfer
Sample
Species
Mus musculus
Sample Type
Cell culture supernatant
Sample Condition
Control condition
EV-producing cells
adipose tissue-derived macrophages
EV-harvesting Medium
EV-depleted serum
Preparation of EDS
Commercial EDS
Isolation Method
Differential ultracentrifugation
dUC: centrifugation steps
Between 800 g and 10,000 g
Between 100,000 g and 150,000 g
Pelleting: time(min)
240-360
Pelleting: rotor type
SW 32 Ti
Pelleting: speed (g)
100000
Pelleting: adjusted k-factor
256.0
Wash: time (min)
20
Wash: Rotor Type
SW 32 Ti
Wash: speed (g)
100000
Wash: adjusted k-factor
256.0
Filtration steps
0.22µm or 0.2µm
EV-subtype
Distinction between multiple subtypes
1.13-1.15 g/ml
Used subtypes
Yes
Characterization: Protein analysis
Protein Concentration Method
DC protein assay
Protein Concentration
9-May
Western Blot
Antibody details provided?
Yes
Lysis buffer provided?
Yes
Detected EV-associated proteins
CD9, CD63, HSP70, TSG101, Syntenin1
Not detected contaminants
Grp94
Fluorescent NTA
Extra information
Full UC protocol not in original article
1 - 2 of 2
  • dUC = differential ultracentrifugation
  • DG = density gradient
  • UF = ultrafiltration
  • SEC = size-exclusion chromatography
EV-TRACK ID
EV170016
species
Mus musculus
sample type
Cell culture
cell type
adipose
tissue-derived macrophages
condition
Obesitas
Control condition
isolation protocol
DG
Filtration
dUC
dUC
Filtration
case number
1
2
EV-METRIC %
88
55