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You searched for: EV200007 (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
EV200007 1/1 Homo sapiens breast milk DG
(d)(U)C
SEC
Zonneveld, Marijke 2021 75%

Study summary

Full title
All authors
Marijke I Zonneveld, Martijn J C van Herwijnen, Marcela M Fernandez-Gutierrez, Alberta Giovanazzi, Anne Marit de Groot, Marije Kleinjan, Toni M M van Capel, Alice J A M Sijts, Leonie S Taams, Johan Garssen, Esther C de Jong, Michiel Kleerebezem, Esther N M Nolte-'t Hoen, Frank A Redegeld, Marca H M Wauben
Journal
J Extracell Vesicles
Abstract
Maternal milk is nature's first functional food. It plays a crucial role in the development of the i (show more...)Maternal milk is nature's first functional food. It plays a crucial role in the development of the infant's gastrointestinal (GI) tract and the immune system. Extracellular vesicles (EVs) are a heterogeneous population of lipid bilayer enclosed vesicles released by cells for intercellular communication and are a component of milk. Recently, we discovered that human milk EVs contain a unique proteome compared to other milk components. Here, we show that physiological concentrations of milk EVs support epithelial barrier function by increasing cell migration via the p38 MAPK pathway. Additionally, milk EVs inhibit agonist-induced activation of endosomal Toll like receptors TLR3 and TLR9. Furthermore, milk EVs directly inhibit activation of CD4+ T cells by temporarily suppressing T cell activation without inducing tolerance. We show that milk EV proteins target key hotspots of signalling networks that can modulate cellular processes in various cell types of the GI tract. (hide)
EV-METRIC
75% (83rd 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
breast milk
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
DG
(d)(U)C
SEC
Protein markers
EV: CD63/ Flotillin1/ CD9/ HSP70
non-EV: Lactoferrin
Proteomics
no
EV density (g/ml)
1.06-1.19
Show all info
Study aim
Function
Sample
Species
Homo sapiens
Sample Type
breast milk
Sample Condition
Control condition
Separation Method
Differential ultracentrifugation
centrifugation steps
Between 800 g and 10,000 g
Between 10,000 g and 50,000 g
Density gradient
Density medium
Iodixanol
Type
Continuous
Lowest density fraction
10%
Highest density fraction
60%
Total gradient volume, incl. sample (mL)
12.5
Sample volume (mL)
6.5
Orientation
Top-down
Rotor type
SW 40 Ti
Speed (g)
192000
Duration (min)
900
Fraction volume (mL)
0.5
Fraction processing
Size-exclusion chromatography
Size-exclusion chromatography
Total column volume (mL)
15
Sample volume/column (mL)
2.5
Resin type
Sephadex G-100
Characterization: Protein analysis
Protein Concentration Method
Not determined
Western Blot
Detected EV-associated proteins
Flotillin1/ CD9/ CD63/ HSP70
Detected contaminants
Lactoferrin
1 - 1 of 1
  • CM = Commercial method
  • dUC = differential ultracentrifugation
  • DG = density gradient
  • UF = ultrafiltration
  • SEC = size-exclusion chromatography
EV-TRACK ID
EV200007
species
Homo sapiens
sample type
breast milk
condition
Control condition
separation protocol
DG
(d)(U)C
SEC
Exp. nr.
1
EV-METRIC %
75