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You searched for: EV160002 (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 and particle analysis
Experiment number
  • Experiments differ in sample type and particle analysis
Experiment number
  • Experiments differ in sample type and particle analysis
Details EV-TRACK ID Experiment nr. Species Sample type Isolation protocol First author Year EV-METRIC
EV160002 2/3 Homo sapiens Extruded cells (NIH3T3) DG
Sequential extrusion
Lunavat TR 2016 37%

Study summary

Full title
All authors
Lunavat TR, Jang SC, Nilsson L, Park HT, Repiska G, Lässer C, Nilsson JA, Gho YS, Lötvall J
Journal
Biomaterials
Abstract
To develop RNA-based therapeutics, it is crucial to create delivery vectors that transport the RNA m (show more...)To develop RNA-based therapeutics, it is crucial to create delivery vectors that transport the RNA molecule into the cell cytoplasm. Naturally released exosomes vesicles (also called "Extracellular Vesicles") have been proposed as possible RNAi carriers, but their yield is relatively small in any cell culture system. We have previously generated exosome-mimetic nanovesicles (NV) by serial extrusions of cells through nano-sized filters, which results in 100-times higher yield of extracellular vesicles. We here test 1) whether NV can be loaded with siRNA exogenously and endogenously, 2) whether the siRNA-loaded NV are taken up by recipient cells, and 3) whether the siRNA can induce functional knock-down responses in recipient cells. A siRNA against GFP was first loaded into NV by electroporation, or a c-Myc shRNA was expressed inside of the cells. The NV were efficiently loaded with siRNA with both techniques, were taken up by recipient cells, which resulted in attenuation of target gene expression. In conclusion, our study suggests that exosome-mimetic nanovesicles can be a platform for RNAi delivery to cell cytoplasm. (hide)
EV-METRIC
37% (50th 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
Extruded cells (NIH3T3)
Focus vesicles
Nanovesicles
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 + Sequential extrusion
Protein markers
EV: PDGFR/ Flotillin-1/ CD9
non-EV: Calnexin
Proteomics
no
Show all info
Study aim
Function, New methodological development
Sample
Species
Homo sapiens
Sample Type
Extruded cells (NIH3T3)
Sample Condition
Control condition
Isolation Method
Density cushion
Density medium
Iodixanol
Other
Name other isolation method
Sequential extrusion
Characterization: Protein analysis
Protein Concentration Method
Bradford
Western Blot
Lysis buffer provided?
Yes
Detected EV-associated proteins
PDGFR, Flotillin-1, CD9
Fluorescent NTA
Characterization: Particle analysis
NTA
Report type
Size range/distribution
Reported size (nm)
180-200
EM
EM-type
Transmission-EM
Image type
Wide-field
EV160002 3/3 Homo sapiens Extruded cells (NIH3T3) DG
Sequential extrusion
Lunavat TR 2016 37%

Study summary

Full title
All authors
Lunavat TR, Jang SC, Nilsson L, Park HT, Repiska G, Lässer C, Nilsson JA, Gho YS, Lötvall J
Journal
Biomaterials
Abstract
To develop RNA-based therapeutics, it is crucial to create delivery vectors that transport the RNA m (show more...)To develop RNA-based therapeutics, it is crucial to create delivery vectors that transport the RNA molecule into the cell cytoplasm. Naturally released exosomes vesicles (also called "Extracellular Vesicles") have been proposed as possible RNAi carriers, but their yield is relatively small in any cell culture system. We have previously generated exosome-mimetic nanovesicles (NV) by serial extrusions of cells through nano-sized filters, which results in 100-times higher yield of extracellular vesicles. We here test 1) whether NV can be loaded with siRNA exogenously and endogenously, 2) whether the siRNA-loaded NV are taken up by recipient cells, and 3) whether the siRNA can induce functional knock-down responses in recipient cells. A siRNA against GFP was first loaded into NV by electroporation, or a c-Myc shRNA was expressed inside of the cells. The NV were efficiently loaded with siRNA with both techniques, were taken up by recipient cells, which resulted in attenuation of target gene expression. In conclusion, our study suggests that exosome-mimetic nanovesicles can be a platform for RNAi delivery to cell cytoplasm. (hide)
EV-METRIC
37% (50th 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
Extruded cells (NIH3T3)
Focus vesicles
Nanovesicles
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 + Sequential extrusion
Protein markers
EV: PDGFR/ Flotillin-1/ CD9
non-EV: Calnexin
Proteomics
no
Show all info
Study aim
Function, New methodological development
Sample
Species
Homo sapiens
Sample Type
Extruded cells (NIH3T3)
Sample Condition
cMyc shRNA
Isolation Method
Density cushion
Density medium
Iodixanol
Other
Name other isolation method
Sequential extrusion
Characterization: Protein analysis
Protein Concentration Method
Bradford
Western Blot
Lysis buffer provided?
Yes
Detected EV-associated proteins
PDGFR, Flotillin-1, CD9
Fluorescent NTA
Characterization: Particle analysis
NTA
Report type
Size range/distribution
Reported size (nm)
180-200
EM
EM-type
Transmission-EM
Image type
Wide-field
EV160002 1/3 Homo sapiens Extruded cells (U973) DG
Sequential extrusion
Lunavat TR 2016 0%

Study summary

Full title
All authors
Lunavat TR, Jang SC, Nilsson L, Park HT, Repiska G, Lässer C, Nilsson JA, Gho YS, Lötvall J
Journal
Biomaterials
Abstract
To develop RNA-based therapeutics, it is crucial to create delivery vectors that transport the RNA m (show more...)To develop RNA-based therapeutics, it is crucial to create delivery vectors that transport the RNA molecule into the cell cytoplasm. Naturally released exosomes vesicles (also called "Extracellular Vesicles") have been proposed as possible RNAi carriers, but their yield is relatively small in any cell culture system. We have previously generated exosome-mimetic nanovesicles (NV) by serial extrusions of cells through nano-sized filters, which results in 100-times higher yield of extracellular vesicles. We here test 1) whether NV can be loaded with siRNA exogenously and endogenously, 2) whether the siRNA-loaded NV are taken up by recipient cells, and 3) whether the siRNA can induce functional knock-down responses in recipient cells. A siRNA against GFP was first loaded into NV by electroporation, or a c-Myc shRNA was expressed inside of the cells. The NV were efficiently loaded with siRNA with both techniques, were taken up by recipient cells, which resulted in attenuation of target gene expression. In conclusion, our study suggests that exosome-mimetic nanovesicles can be a platform for RNAi delivery to cell cytoplasm. (hide)
EV-METRIC
0% (median: 0% 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
Extruded cells (U973)
Focus vesicles
Nanovesicles
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 + Sequential extrusion
Protein markers
EV: None
non-EV: None
Proteomics
no
Show all info
Study aim
Function, New methodological development
Sample
Species
Homo sapiens
Sample Type
Extruded cells (U973)
Sample Condition
Control condition
Isolation Method
Density cushion
Density medium
Iodixanol
Other
Name other isolation method
Sequential extrusion
Protein Concentration Method
Bradford
Fluorescent NTA
Characterization: Particle analysis
DLS
Report type
Modus
Reported size (nm)
150
1 - 3 of 3
  • dUC = differential ultracentrifugation
  • DG = density gradient
  • UF = ultrafiltration
  • SEC = size-exclusion chromatography
EV-TRACK ID
EV160002
species
Homo sapiens
sample type
Extruded
cells (NIH3T3)
Extruded
cells (NIH3T3)
Extruded
cells (U973)
condition
Control condition
cMyc shRNA
Control condition
isolation protocol
DG
Sequential extrusion
DG
Sequential extrusion
DG
Sequential extrusion
case number
2
3
1
EV-METRIC %
37
37
0