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Details	EV-TRACK ID	Experiment nr.	Species	Sample type	Separation protocol	First author	Year	EV-METRIC
	EV170013	1/1	Mus musculus	endothelial progenitor cells	DG (d)(U)C	Degosserie, Jonathan	2018	100%
Study summary Full title Extracellular vesicles from endothelial progenitor cells promote thyroid follicle formation. All authors Degosserie J, Heymans C, Spourquet C, Halbout M, D'Auria L, Van Der Smissen P, Vertommen D, Courtoy PJ, Tyteca D, Pierreux CE. Journal J Extracell Vesicles Abstract Organogenesis is a complex and dynamic process requiring reciprocal communication between different (show more...)Organogenesis is a complex and dynamic process requiring reciprocal communication between different cell types. In the thyroid, thyrocyte progenitors secrete the angiocrine factor, VEGFA, to recruit endothelial cells. In return, endothelial cells promote thyrocyte organisation into spherical follicular structures, which are responsible for thyroid hormone synthesis and storage. Medium conditioned by endothelial progenitor cells (EPCs) can promote follicle formation and lumen expansion (i.e. folliculogenesis) in an ex vivo culture system of thyroid lobes. Here, we postulated that endothelial cells instruct thyrocyte progenitors by producing extracellular vesicles (EVs). We found that medium conditioned by EPCs contain EVs with exosomal characteristics and that these vesicles can be incorporated into thyrocyte progenitors. By mass spectrometry, laminin peptides were abundantly identified in the EV preparations, probably co-sedimenting with EVs. Laminin-α1 silencing in EPC abrogated the folliculogenic effect of EVs. However, density gradient separation of EVs from laminins revealed that both EV-rich and laminin-rich fractions exhibited folliculogenic activity. In conclusion, we suggest that endothelial cells can produce EVs favouring thyrocyte organisation into follicles and lumen expansion, a mechanism promoted by laminin-α1. (hide) EV-METRIC 100% (99th 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 DG (d)(U)C Adj. k-factor 84.53 (pelleting) / 84.53 (washing) Protein markers EV: Flotillin-1/ CD63/ CD9 non-EV: Calnexin Proteomics yes Show all info Study aim Function, Identification of content (omics approaches) Sample Species Mus musculus Sample Type Cell culture supernatant EV-producing cells endothelial progenitor cells EV-harvesting Medium Serum free medium Separation Method (Differential) (ultra)centrifugation dUC: centrifugation steps Between 800 g and 10,000 g Between 10,000 g and 50,000 g Equal to or above 150,000 g Pelleting performed Yes Pelleting: time(min) 90 Pelleting: rotor type Type 80 Ti Pelleting: speed (g) 150000 Pelleting: adjusted k-factor 84.53 Wash: time (min) 90 Wash: Rotor Type Type 80 Ti Wash: speed (g) 150000 Wash: adjusted k-factor 84.53 Density gradient Only used for validation of main results Yes Type Discontinuous Number of initial discontinuous layers 3 Lowest density fraction 10% Highest density fraction 30% Sample volume (mL) 1.3 Orientation Bottom-up (sample migrates upwards) Rotor type SW 55 Ti Speed (g) 100000 Duration (min) 960 Fraction volume (mL) 0.625 Fraction processing Centrifugation Pelleting: volume per fraction 8 Pelleting: duration (min) 90 Pelleting: rotor type Type 80 Ti Pelleting: speed (g) 150000 Pelleting: adjusted k-factor 84.53 Characterization: Protein analysis Protein Concentration Method BCA Western Blot Detected EV-associated proteins CD9, CD63, Flotillin-1 Not detected contaminants Calnexin Proteomics database No Characterization: Lipid analysis No Characterization: Particle analysis NTA Report type Size range/distribution Reported size (nm) 100-150 EV concentration Yes Particle yield 3.60E+08 particles/million cells EM EM-type Scanning-EM Image type Close-up, Wide-field

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EV-TRACK ID	EV170013
species	Mus musculus
sample type	Cell culture
cell type	endothelial progenitor cells
condition	Control condition
separation protocol	DG (d)(U)C
Exp. nr.	1
EV-METRIC %	100