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You searched for: EV170012 (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
Experiment number
  • Experiments differ in Sample type
Experiment number
  • Experiments differ in Sample type
Experiment number
  • Experiments differ in Sample type
Details EV-TRACK ID Experiment nr. Species Sample type separation protocol First author Year EV-METRIC
EV170012 1/3 Homo sapiens Cell culture supernatant (d)(U)C Gualerzi, Alice 2017 56%

Study summary

Full title
All authors
Alice Gualerzi, Stefania Niada, Chiara Giannasi, Silvia Picciolini, Carlo Morasso, Renzo Vanna, Valeria Rossella, Massimo Masserini, Marzia Bedoni, Fabio Ciceri, Maria Ester Bernardo, Anna Teresa Brini & Furio Gramatica
Journal
Scientific Reports
Abstract
Extracellular vesicles (EVs) from mesenchymal stromal cells (MSC) are emerging as valuable therapeut (show more...)Extracellular vesicles (EVs) from mesenchymal stromal cells (MSC) are emerging as valuable therapeutic agents for tissue regeneration and immunomodulation, but their clinical applications have so far been limited by the technical restraints of current isolation and characterisation procedures. This study shows for the first time the successful application of Raman spectroscopy as label-free, sensitive and reproducible means of carrying out the routine bulk characterisation of MSC-derived vesicles before their use in vitro or in vivo, thus promoting the translation of EV research to clinical practice. The Raman spectra of the EVs of bone marrow and adipose tissue-derived MSCs were compared with human dermal fibroblast EVs in order to demonstrate the ability of the method to distinguish the vesicles of the three cytotypes automatically with an accuracy of 93.7%. Our data attribute a Raman fingerprint to EVs from undifferentiated and differentiated cells of diverse tissue origin, and provide insights into the biochemical characteristics of EVs from different sources and into the differential contribution of sphingomyelin, gangliosides and phosphatidilcholine to the Raman spectra themselves. (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
adipose tissue mesenchymal stromal cells
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
Adj. k-factor
158.5 (pelleting) / 158.5 (washing)
Protein markers
EV: Flotillin-1/ CD63/ CD9
non-EV: Calnexin
Proteomics
yes
Show all info
Study aim
New methodological development
Sample
Species
Homo sapiens
Sample Type
Cell culture supernatant
Sample Condition
Control condition
EV-producing cells
adipose tissue mesenchymal stromal cells
EV-harvesting Medium
Serum free medium
Separation Method
Differential ultracentrifugation
centrifugation steps
Below or equal to 800 g
Between 800 g and 10,000 g
Between 100,000 g and 150,000 g
Obtain an EV pellet :
Yes
Pelleting: time(min)
70
Pelleting: rotor type
Type 55.2 Ti
Pelleting: speed (g)
100000
Pelleting: adjusted k-factor
158.5
Wash: time (min)
70
Wash: Rotor Type
Type 55.2 Ti
Wash: speed (g)
100000
Wash: adjusted k-factor
158.5
Characterization: Protein analysis
Protein Concentration Method
BCA
Western Blot
Lysis buffer provided?
Yes
Detected EV-associated proteins
CD9, CD63, Flotillin-1
Not detected contaminants
Calnexin
Characterization: Particle analysis
EM
EM-type
Transmission-EM
Image type
Wide-field
Report size (nm)
46.5 ± 15.8
Other particle analysis name(1)
Raman spectroscopy
Extra information
Primary antibodies: - Purified Mouse Anti-Flotillin-1; Clone 18/Flotillin-1; BD Transduction Laboratories™, San Jose, CA, USA - Rabbit anti-CD63; System Biosciences, Palo Alto, CA, USA - Rabbit anti-CD9; System Biosciences, Palo Alto, CA, USA - Rabbit anti-calnexin; clone C5C9, Cell Signaling Technology, Danvers, MA, USA
EV170012 2/3 Homo sapiens Cell culture supernatant (d)(U)C Gualerzi, Alice 2017 56%

Study summary

Full title
All authors
Alice Gualerzi, Stefania Niada, Chiara Giannasi, Silvia Picciolini, Carlo Morasso, Renzo Vanna, Valeria Rossella, Massimo Masserini, Marzia Bedoni, Fabio Ciceri, Maria Ester Bernardo, Anna Teresa Brini & Furio Gramatica
Journal
Scientific Reports
Abstract
Extracellular vesicles (EVs) from mesenchymal stromal cells (MSC) are emerging as valuable therapeut (show more...)Extracellular vesicles (EVs) from mesenchymal stromal cells (MSC) are emerging as valuable therapeutic agents for tissue regeneration and immunomodulation, but their clinical applications have so far been limited by the technical restraints of current isolation and characterisation procedures. This study shows for the first time the successful application of Raman spectroscopy as label-free, sensitive and reproducible means of carrying out the routine bulk characterisation of MSC-derived vesicles before their use in vitro or in vivo, thus promoting the translation of EV research to clinical practice. The Raman spectra of the EVs of bone marrow and adipose tissue-derived MSCs were compared with human dermal fibroblast EVs in order to demonstrate the ability of the method to distinguish the vesicles of the three cytotypes automatically with an accuracy of 93.7%. Our data attribute a Raman fingerprint to EVs from undifferentiated and differentiated cells of diverse tissue origin, and provide insights into the biochemical characteristics of EVs from different sources and into the differential contribution of sphingomyelin, gangliosides and phosphatidilcholine to the Raman spectra themselves. (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
dermal fibroblasts
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
Adj. k-factor
158.5 (pelleting) / 158.5 (washing)
Protein markers
EV: Flotillin-1/ CD63/ CD9
non-EV: Calnexin
Proteomics
yes
Show all info
Study aim
New methodological development
Sample
Species
Homo sapiens
Sample Type
Cell culture supernatant
Sample Condition
Control condition
EV-producing cells
dermal fibroblasts
EV-harvesting Medium
Serum free medium
Separation Method
Differential ultracentrifugation
centrifugation steps
Below or equal to 800 g
Between 800 g and 10,000 g
Between 100,000 g and 150,000 g
Obtain an EV pellet :
Yes
Pelleting: time(min)
70
Pelleting: rotor type
Type 55.2 Ti
Pelleting: speed (g)
100000
Pelleting: adjusted k-factor
158.5
Wash: time (min)
70
Wash: Rotor Type
Type 55.2 Ti
Wash: speed (g)
100000
Wash: adjusted k-factor
158.5
Characterization: Protein analysis
Protein Concentration Method
BCA
Western Blot
Lysis buffer provided?
Yes
Detected EV-associated proteins
CD9, CD63, Flotillin-1
Not detected contaminants
Calnexin
Characterization: Particle analysis
EM
EM-type
Transmission-EM
Image type
Wide-field
Report size (nm)
46.5 ± 15.8
Other particle analysis name(1)
Raman spectroscopy
Extra information
Primary antibodies: - Purified Mouse Anti-Flotillin-1; Clone 18/Flotillin-1; BD Transduction Laboratories™, San Jose, CA, USA - Rabbit anti-CD63; System Biosciences, Palo Alto, CA, USA - Rabbit anti-CD9; System Biosciences, Palo Alto, CA, USA - Rabbit anti-calnexin; clone C5C9, Cell Signaling Technology, Danvers, MA, USA
EV170012 3/3 Homo sapiens Cell culture supernatant (d)(U)C Gualerzi, Alice 2017 56%

Study summary

Full title
All authors
Alice Gualerzi, Stefania Niada, Chiara Giannasi, Silvia Picciolini, Carlo Morasso, Renzo Vanna, Valeria Rossella, Massimo Masserini, Marzia Bedoni, Fabio Ciceri, Maria Ester Bernardo, Anna Teresa Brini & Furio Gramatica
Journal
Scientific Reports
Abstract
Extracellular vesicles (EVs) from mesenchymal stromal cells (MSC) are emerging as valuable therapeut (show more...)Extracellular vesicles (EVs) from mesenchymal stromal cells (MSC) are emerging as valuable therapeutic agents for tissue regeneration and immunomodulation, but their clinical applications have so far been limited by the technical restraints of current isolation and characterisation procedures. This study shows for the first time the successful application of Raman spectroscopy as label-free, sensitive and reproducible means of carrying out the routine bulk characterisation of MSC-derived vesicles before their use in vitro or in vivo, thus promoting the translation of EV research to clinical practice. The Raman spectra of the EVs of bone marrow and adipose tissue-derived MSCs were compared with human dermal fibroblast EVs in order to demonstrate the ability of the method to distinguish the vesicles of the three cytotypes automatically with an accuracy of 93.7%. Our data attribute a Raman fingerprint to EVs from undifferentiated and differentiated cells of diverse tissue origin, and provide insights into the biochemical characteristics of EVs from different sources and into the differential contribution of sphingomyelin, gangliosides and phosphatidilcholine to the Raman spectra themselves. (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
bone marrow-derived mesenchymal stromal cells
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
Adj. k-factor
158.5 (pelleting) / 158.5 (washing)
Protein markers
EV: Flotillin-1/ CD63/ CD9
non-EV: Calnexin
Proteomics
no
Show all info
Study aim
New methodological development
Sample
Species
Homo sapiens
Sample Type
Cell culture supernatant
Sample Condition
Control condition
EV-producing cells
bone marrow-derived mesenchymal stromal cells
EV-harvesting Medium
Serum free medium
Separation Method
Differential ultracentrifugation
centrifugation steps
Below or equal to 800 g
Between 800 g and 10,000 g
Between 100,000 g and 150,000 g
Obtain an EV pellet :
Yes
Pelleting: time(min)
70
Pelleting: rotor type
Type 55.2 Ti
Pelleting: speed (g)
100000
Pelleting: adjusted k-factor
158.5
Wash: time (min)
70
Wash: Rotor Type
Type 55.2 Ti
Wash: speed (g)
100000
Wash: adjusted k-factor
158.5
Characterization: Protein analysis
Protein Concentration Method
BCA
Western Blot
Lysis buffer provided?
Yes
Detected EV-associated proteins
CD9, CD63, Flotillin-1
Not detected contaminants
Calnexin
Characterization: Particle analysis
EM
EM-type
Transmission-EM
Image type
Wide-field
Report size (nm)
46.5 ± 15.8
Other particle analysis name(1)
Raman spectroscopy
Extra information
Primary antibodies: - Purified Mouse Anti-Flotillin-1; Clone 18/Flotillin-1; BD Transduction Laboratories™, San Jose, CA, USA - Rabbit anti-CD63; System Biosciences, Palo Alto, CA, USA - Rabbit anti-CD9; System Biosciences, Palo Alto, CA, USA - Rabbit anti-calnexin; clone C5C9, Cell Signaling Technology, Danvers, MA, USA
1 - 3 of 3
  • CM = Commercial method
  • dUC = differential ultracentrifugation
  • DG = density gradient
  • UF = ultrafiltration
  • SEC = size-exclusion chromatography
EV-TRACK ID
EV170012
species
Homo sapiens
sample type
Cell culture
cell type
adipose
tissue
mesenchymal stromal
cells
dermal fibroblasts
bone
marrow-derived
mesenchymal stromal
cells
condition
Control condition
Control condition
Control condition
separation protocol
(d)(U)C
(d)(U)C
(d)(U)C
Exp. nr.
1
2
3
EV-METRIC %
56
56
56