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You searched for: EV200014 (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
    • IAF = immuno-affinity capture
Details EV-TRACK ID Experiment nr. Species Sample type separation protocol First author Year EV-METRIC
EV200014 1/4 Homo sapiens Cell culture supernatant (d)(U)C
qEV
Wallis, Ryan 2021 56%

Study summary

Full title
All authors
Ryan Wallis, Natasa Josipovic, Hannah Mizen, Arturo Robles‐Tenorio, Eleanor J. Tyler, Argyris Papantonis, Cleo L. Bishop
Journal
J Extracell Vesicles
Abstract
A hallmark of senescence is the acquisition of an enhanced secretome comprising inflammatory mediato (show more...)A hallmark of senescence is the acquisition of an enhanced secretome comprising inflammatory mediators and tissue remodelling agents – the senescence‐associated secretory phenotype (SASP). Through the SASP, senescent cells are hypothesised to contribute to both ageing and pathologies associated with age. Whilst soluble factors have been the most widely investigated components of the SASP, there is growing evidence that small extracellular vesicles (EVs) comprise functionally important constituents. Thus, dissecting the contribution of the soluble SASP from the vesicular component is crucial to elucidating the functional significance of senescent cell derived EVs. Here, we take advantage of a systematic proteomics based approach to determine that soluble SASP factors co‐isolate with EVs following differential ultracentrifugation (dUC). We present size‐exclusion chromatography (SEC) as a method for separation of the soluble and vesicular components of the senescent secretome and thus EV purification. Furthermore, we demonstrate that SEC EVs isolated from senescent cells contribute to non‐cell autonomous paracrine senescence. Therefore, this work emphasises the requirement for methodological rigor due to the propensity of SASP components to co‐isolate during dUC and provides a framework for future investigations of the vesicular component of the SASP. (hide)
EV-METRIC
56% (84th 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
Cell Name
IMR90 ER:STOP
Sample origin
Control condition
Focus vesicles
extracellular vesicle
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
    • IAF = immuno-affinity capture
(d)(U)C
qEV
Protein markers
EV: TSG101/ CD9
non-EV: Calnexin
Proteomics
no
Show all info
Study aim
Function/Identification of content (omics approaches)/Technical analysis comparing/optimizing EV-related methods
Sample
Species
Homo sapiens
Sample Type
Cell culture supernatant
Sample Condition
Control condition
EV-producing cells
IMR90 ER:STOP
EV-harvesting Medium
EV-depleted medium
Preparation of EDS
Commercial EDS
Cell number specification
No
Separation Method
Differential ultracentrifugation
centrifugation steps
Between 800 g and 10,000 g
Between 10,000 g and 50,000 g
Between 100,000 g and 150,000 g
Obtain an EV pellet :
Yes
Pelleting: time(min)
90
Pelleting: rotor type
T-865
Pelleting: speed (g)
99582
Commercial kit
qEV
Characterization: Protein analysis
PMID previous EV protein analysis
Extra characterization
Protein Concentration Method
microBCA
Western Blot
Not detected EV-associated proteins
TSG101/ CD9
Not detected contaminants
Calnexin
Characterization: Particle analysis
PMID previous EV particle analysis
Extra particle analysis
NTA
Report type
Modus
Reported size (nm)
92
EV concentration
Yes
Particle yield
total concentration;Yes, other: 1.60E+10
EV200014 2/4 Homo sapiens Cell culture supernatant (d)(U)C Wallis, Ryan 2021 56%

Study summary

Full title
All authors
Ryan Wallis, Natasa Josipovic, Hannah Mizen, Arturo Robles‐Tenorio, Eleanor J. Tyler, Argyris Papantonis, Cleo L. Bishop
Journal
J Extracell Vesicles
Abstract
A hallmark of senescence is the acquisition of an enhanced secretome comprising inflammatory mediato (show more...)A hallmark of senescence is the acquisition of an enhanced secretome comprising inflammatory mediators and tissue remodelling agents – the senescence‐associated secretory phenotype (SASP). Through the SASP, senescent cells are hypothesised to contribute to both ageing and pathologies associated with age. Whilst soluble factors have been the most widely investigated components of the SASP, there is growing evidence that small extracellular vesicles (EVs) comprise functionally important constituents. Thus, dissecting the contribution of the soluble SASP from the vesicular component is crucial to elucidating the functional significance of senescent cell derived EVs. Here, we take advantage of a systematic proteomics based approach to determine that soluble SASP factors co‐isolate with EVs following differential ultracentrifugation (dUC). We present size‐exclusion chromatography (SEC) as a method for separation of the soluble and vesicular components of the senescent secretome and thus EV purification. Furthermore, we demonstrate that SEC EVs isolated from senescent cells contribute to non‐cell autonomous paracrine senescence. Therefore, this work emphasises the requirement for methodological rigor due to the propensity of SASP components to co‐isolate during dUC and provides a framework for future investigations of the vesicular component of the SASP. (hide)
EV-METRIC
56% (84th 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
Cell Name
IMR90 ER:STOP
Sample origin
Control condition
Focus vesicles
extracellular vesicle
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
    • IAF = immuno-affinity capture
(d)(U)C
Protein markers
EV: TSG101/ IL-8/ CD9
non-EV: Calnexin
Proteomics
yes
Show all info
Study aim
Function/Identification of content (omics approaches)/Technical analysis comparing/optimizing EV-related methods
Sample
Species
Homo sapiens
Sample Type
Cell culture supernatant
Sample Condition
Control condition
EV-producing cells
IMR90 ER:STOP
EV-harvesting Medium
EV-depleted medium
Preparation of EDS
Commercial EDS
Cell number specification
No
Separation Method
Differential ultracentrifugation
centrifugation steps
Between 800 g and 10,000 g
Between 10,000 g and 50,000 g
Between 100,000 g and 150,000 g
Obtain an EV pellet :
Yes
Pelleting: time(min)
90
Pelleting: rotor type
T-865
Pelleting: speed (g)
99582
Characterization: Protein analysis
PMID previous EV protein analysis
Extra characterization
Protein Concentration Method
Not determined
Western Blot
Detected EV-associated proteins
CD9/ TSG101
Detected contaminants
Calnexin
ELISA
Detected EV-associated proteins
IL-8
Proteomics
Proteomics database
No
Characterization: Particle analysis
PMID previous EV particle analysis
Extra particle analysis
NTA
Report type
Modus
Reported size (nm)
85.2
EV concentration
Yes
Particle yield
total concentration;Yes, other: 2.20E+11
EV200014 3/4 Homo sapiens Cell culture supernatant (d)(U)C
qEV
Wallis, Ryan 2021 56%

Study summary

Full title
All authors
Ryan Wallis, Natasa Josipovic, Hannah Mizen, Arturo Robles‐Tenorio, Eleanor J. Tyler, Argyris Papantonis, Cleo L. Bishop
Journal
J Extracell Vesicles
Abstract
A hallmark of senescence is the acquisition of an enhanced secretome comprising inflammatory mediato (show more...)A hallmark of senescence is the acquisition of an enhanced secretome comprising inflammatory mediators and tissue remodelling agents – the senescence‐associated secretory phenotype (SASP). Through the SASP, senescent cells are hypothesised to contribute to both ageing and pathologies associated with age. Whilst soluble factors have been the most widely investigated components of the SASP, there is growing evidence that small extracellular vesicles (EVs) comprise functionally important constituents. Thus, dissecting the contribution of the soluble SASP from the vesicular component is crucial to elucidating the functional significance of senescent cell derived EVs. Here, we take advantage of a systematic proteomics based approach to determine that soluble SASP factors co‐isolate with EVs following differential ultracentrifugation (dUC). We present size‐exclusion chromatography (SEC) as a method for separation of the soluble and vesicular components of the senescent secretome and thus EV purification. Furthermore, we demonstrate that SEC EVs isolated from senescent cells contribute to non‐cell autonomous paracrine senescence. Therefore, this work emphasises the requirement for methodological rigor due to the propensity of SASP components to co‐isolate during dUC and provides a framework for future investigations of the vesicular component of the SASP. (hide)
EV-METRIC
56% (84th 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
Cell Name
IMR90 ER:RAS
Sample origin
HRas:ER Oncogene Induced Senescence Model
Focus vesicles
extracellular vesicle
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
    • IAF = immuno-affinity capture
(d)(U)C
qEV
Protein markers
EV: TSG101/ IL-8/ CD9
non-EV: Calnexin
Proteomics
yes
Show all info
Study aim
Function/Identification of content (omics approaches)/Technical analysis comparing/optimizing EV-related methods
Sample
Species
Homo sapiens
Sample Type
Cell culture supernatant
Sample Condition
HRas:ER Oncogene Induced Senescence Model
EV-producing cells
IMR90 ER:RAS
EV-harvesting Medium
EV-depleted medium
Preparation of EDS
Commercial EDS
Cell number specification
No
Separation Method
Differential ultracentrifugation
centrifugation steps
Between 800 g and 10,000 g
Between 10,000 g and 50,000 g
Between 100,000 g and 150,000 g
Obtain an EV pellet :
Yes
Pelleting: time(min)
90
Pelleting: rotor type
T-865
Pelleting: speed (g)
99582
Commercial kit
qEV
Characterization: Protein analysis
PMID previous EV protein analysis
Extra characterization
Protein Concentration Method
microBCA
Western Blot
Detected EV-associated proteins
CD9/ TSG101
Not detected contaminants
Calnexin
ELISA
Detected EV-associated proteins
IL-8
Proteomics
Proteomics database
No
Characterization: Particle analysis
PMID previous EV particle analysis
Extra particle analysis
NTA
Report type
Modus
Reported size (nm)
107
EV concentration
Yes
Particle yield
total concentration;Yes, other: 8.80E+10
EV200014 4/4 Homo sapiens Cell culture supernatant (d)(U)C Wallis, Ryan 2021 56%

Study summary

Full title
All authors
Ryan Wallis, Natasa Josipovic, Hannah Mizen, Arturo Robles‐Tenorio, Eleanor J. Tyler, Argyris Papantonis, Cleo L. Bishop
Journal
J Extracell Vesicles
Abstract
A hallmark of senescence is the acquisition of an enhanced secretome comprising inflammatory mediato (show more...)A hallmark of senescence is the acquisition of an enhanced secretome comprising inflammatory mediators and tissue remodelling agents – the senescence‐associated secretory phenotype (SASP). Through the SASP, senescent cells are hypothesised to contribute to both ageing and pathologies associated with age. Whilst soluble factors have been the most widely investigated components of the SASP, there is growing evidence that small extracellular vesicles (EVs) comprise functionally important constituents. Thus, dissecting the contribution of the soluble SASP from the vesicular component is crucial to elucidating the functional significance of senescent cell derived EVs. Here, we take advantage of a systematic proteomics based approach to determine that soluble SASP factors co‐isolate with EVs following differential ultracentrifugation (dUC). We present size‐exclusion chromatography (SEC) as a method for separation of the soluble and vesicular components of the senescent secretome and thus EV purification. Furthermore, we demonstrate that SEC EVs isolated from senescent cells contribute to non‐cell autonomous paracrine senescence. Therefore, this work emphasises the requirement for methodological rigor due to the propensity of SASP components to co‐isolate during dUC and provides a framework for future investigations of the vesicular component of the SASP. (hide)
EV-METRIC
56% (84th 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
Cell Name
IMR90 ER:RAS
Sample origin
HRas:ER Oncogene Induced Senescence Model
Focus vesicles
extracellular vesicle
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
    • IAF = immuno-affinity capture
(d)(U)C
Protein markers
EV: TSG101/ IL-8/ CD9
non-EV: Calnexin
Proteomics
yes
Show all info
Study aim
Function/Identification of content (omics approaches)/Technical analysis comparing/optimizing EV-related methods
Sample
Species
Homo sapiens
Sample Type
Cell culture supernatant
Sample Condition
HRas:ER Oncogene Induced Senescence Model
EV-producing cells
IMR90 ER:RAS
EV-harvesting Medium
EV-depleted medium
Preparation of EDS
Commercial EDS
Cell number specification
No
Separation Method
Differential ultracentrifugation
centrifugation steps
Between 800 g and 10,000 g
Between 10,000 g and 50,000 g
Between 100,000 g and 150,000 g
Obtain an EV pellet :
Yes
Pelleting: time(min)
90
Pelleting: rotor type
T-865
Pelleting: speed (g)
99582
Characterization: Protein analysis
PMID previous EV protein analysis
Extra characterization
Protein Concentration Method
Not determined
Western Blot
Detected EV-associated proteins
CD9/ TSG101
Detected contaminants
Calnexin
ELISA
Detected EV-associated proteins
IL-8
Proteomics
Proteomics database
No
Characterization: Particle analysis
PMID previous EV particle analysis
Extra particle analysis
NTA
Report type
Modus
Reported size (nm)
102
EV concentration
Yes
Particle yield
total concentration;Yes, other: 4.50E+11
1 - 4 of 4
  • CM = Commercial method
  • dUC = differential ultracentrifugation
  • DG = density gradient
  • UF = ultrafiltration
  • SEC = size-exclusion chromatography
EV-TRACK ID
EV200014
species
Homo sapiens
sample type
Cell culture
cell type
IMR90 ER:STOP
IMR90 ER:STOP
IMR90 ER:RAS
IMR90 ER:RAS
condition
Control condition
Control condition
HRas:ER
Oncogene Induced
Senescence Model
HRas:ER
Oncogene Induced
Senescence Model
separation protocol
(d)(U)C
qEV
(d)(U)C
(d)(U)C
qEV
(d)(U)C
Exp. nr.
1
2
3
4
EV-METRIC %
56
56
56
56