EV-TRACK

Search > Results

You searched for: EV180024 (EV-TRACK ID)

Showing 1 - 2 of 2

Results list Study Tree

Details	EV-TRACK ID	Experiment nr.	Species	Sample type	Separation protocol	First author	Year	EV-METRIC
	EV180024	1/2	Homo sapiens	THP-1	(d)(U)C Filtration	Rice TF	2018	55%
Study summary Full title Macrophage- but not monocyte-derived extracellular vesicles induce placental pro-inflammatory responses. All authors Rice TF, Donaldson B, Bouqueau M, Kampmann B, Holder B. Journal Placenta Abstract The placenta sheds extracellular vesicles (EVs), including exosomes, into the maternal circulation. (show more...)The placenta sheds extracellular vesicles (EVs), including exosomes, into the maternal circulation. We recently demonstrated that this trafficking of EVs is bi-directional; with uptake of macrophage exosomes by the placenta inducing cytokine release. The specificity of this response is currently unknown. THP-1 cells were cultured as monocytes or differentiated to macrophages, and EVs isolated by ultra-centrifugation. The effect of EVs on human placental explants was measured by cytokine ELISA/luminex. Macrophage, but not monocyte, EVs induce the release of pro-inflammatory cytokines by the placenta. Thus, placental responses to immune cell EVs, including exosomes, reflects the phenotype of the source cell. (hide) EV-METRIC 55% (88th 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. For the quantitative method, the reporting of measured EV concentration is expected. 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 Sample origin PMA-stimulated Focus vesicles extracellular vesicle Separation protocol Separation protocol Gives a short, non-chronological overview of the different steps of the separation protocol. dUC = (Differential) (ultra)centrifugation DG = density gradient UF = ultrafiltration SEC = size-exclusion chromatography IAF = immuno-affinity capture (d)(U)C Filtration Adj. k-factor 228.8 (pelleting) Protein markers EV: Flotillin-1/ MHC1/ TSG101 non-EV: Calnexin Proteomics no Show all info Study aim Function Sample Species Homo sapiens Sample Type Cell culture supernatant EV-producing cells THP-1 EV-harvesting Medium EV-depleted serum Preparation of EDS Commercial EDS Separation Method (Differential) (ultra)centrifugation dUC: 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 Pelleting performed Yes Pelleting: time(min) 90 Pelleting: rotor type F-28/50 Pelleting: speed (g) 100000 Pelleting: adjusted k-factor 228.8 Filtration steps 0.22µm or 0.2µm Characterization: Protein analysis Protein Concentration Method BCA Protein Yield (µg) 3.24 Western Blot Antibody details provided? Yes Lysis buffer provided? Yes Detected EV-associated proteins Flotillin-1, MHC1, TSG101 Not detected contaminants Calnexin Characterization: Lipid analysis No Characterization: Particle analysis NTA Report type Modus Reported size (nm) 111 EV concentration Yes Particle yield 1400000000 Extra information Cells were checked for viability visually, but as they are adherent and impossible to get off the plate in good numbers whilst preserving viability, it is not possible to report viability accurately at time of EV collection. Both mode and median size were reported.

	EV180024	2/2	Homo sapiens	THP-1	(d)(U)C Filtration	Rice TF	2018	55%
Study summary Full title Macrophage- but not monocyte-derived extracellular vesicles induce placental pro-inflammatory responses. All authors Rice TF, Donaldson B, Bouqueau M, Kampmann B, Holder B. Journal Placenta Abstract The placenta sheds extracellular vesicles (EVs), including exosomes, into the maternal circulation. (show more...)The placenta sheds extracellular vesicles (EVs), including exosomes, into the maternal circulation. We recently demonstrated that this trafficking of EVs is bi-directional; with uptake of macrophage exosomes by the placenta inducing cytokine release. The specificity of this response is currently unknown. THP-1 cells were cultured as monocytes or differentiated to macrophages, and EVs isolated by ultra-centrifugation. The effect of EVs on human placental explants was measured by cytokine ELISA/luminex. Macrophage, but not monocyte, EVs induce the release of pro-inflammatory cytokines by the placenta. Thus, placental responses to immune cell EVs, including exosomes, reflects the phenotype of the source cell. (hide) EV-METRIC 55% (88th 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. For the quantitative method, the reporting of measured EV concentration is expected. 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 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. dUC = (Differential) (ultra)centrifugation DG = density gradient UF = ultrafiltration SEC = size-exclusion chromatography IAF = immuno-affinity capture (d)(U)C Filtration Adj. k-factor 228.8 (pelleting) Protein markers EV: Flotillin-1/ MHC1/ TSG101 non-EV: Calnexin Proteomics no Show all info Study aim Function Sample Species Homo sapiens Sample Type Cell culture supernatant EV-producing cells THP-1 EV-harvesting Medium EV-depleted serum Preparation of EDS Commercial EDS Separation Method (Differential) (ultra)centrifugation dUC: 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 Pelleting performed Yes Pelleting: time(min) 90 Pelleting: rotor type F-28/50 Pelleting: speed (g) 100000 Pelleting: adjusted k-factor 228.8 Filtration steps 0.22µm or 0.2µm Characterization: Protein analysis Protein Concentration Method BCA Protein Yield (µg) 8.44 Western Blot Antibody details provided? Yes Lysis buffer provided? Yes Detected EV-associated proteins Flotillin-1, MHC1, TSG101 Not detected contaminants Calnexin Characterization: Lipid analysis No Characterization: Particle analysis NTA Report type Modus Reported size (nm) 86 EV concentration Yes Particle yield 3200000000 Extra information Cells were checked for viability visually, but as they are adherent and impossible to get off the plate in good numbers whilst preserving viability, it is not possible to report viability accurately at time of EV collection. Both mode and median size were reported.

				1 - 2 of 2

EV-TRACK ID	EV180024
species	Homo sapiens
sample type	Cell culture
cell type	THP-1
condition	PMA-stimulated	Control condition
separation protocol	(d)(U)C Filtration	(d)(U)C Filtration
Exp. nr.	1	2
EV-METRIC %	55	55