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Details	EV-TRACK ID	Experiment nr.	Species	Sample type	Separation protocol	First author	Year	EV-METRIC
	EV230000	1/2	Bos taurus	Bovine Oviductal Epithelial Cell Organoids	(d)(U)C Filtration Size-exclusion chromatography (non-commercial)	Menjivar NG	2023	67%
Study summary Full title Bovine oviductal organoids: a multi-omics approach to capture the cellular and extracellular molecular response of the oviduct to heat stress. All authors Menjivar NG, Gad A, Thompson RE, Meyers MA, Hollinshead FK, Tesfaye D Journal BMC Genomics Abstract The mammalian oviduct is a complex, fibromuscular organ known for its role in orchestrating a series (show more...)The mammalian oviduct is a complex, fibromuscular organ known for its role in orchestrating a series of timely and dynamic changes to suitably support early embryogenesis. Climate change-induced heat stress (HS) is one of the largest single stressors compromising reproductive function in humans and farm animals via systemic changes in the redox status of the maternal environment, adversely affecting fertilization and early embryonic development. Oviductal organoids represent a unique 3-dimensional, biomimetic model to study the physiology of the oviduct and its subsequent impact on embryo development under various environmental conditions. (hide) EV-METRIC 67% (94th 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 (Differential) (ultra)centrifugation Filtration Size-exclusion chromatography (non-commercial) Protein markers EV: CD63/ FLOT1/ TSG101 non-EV: Cytochrome C Proteomics no Show all info Study aim Identification of content (omics approaches) Sample Species Bos taurus Sample Type Cell culture supernatant EV-producing cells Bovine Oviductal Epithelial Cell Organoids EV-harvesting Medium Serum free medium Separation Method (Differential) (ultra)centrifugation dUC: centrifugation steps Below or equal to 800 g 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: rotor type SW 55 Ti Pelleting: speed (g) 120000 Wash: volume per pellet (ml) 3 Wash: time (min) 70 Wash: Rotor Type SW 55 Ti Wash: speed (g) 120000 Filtration steps 0.2 or 0.22 µm Size-exclusion chromatography Total column volume (mL) 0.1 Sample volume/column (mL) 0.1 Other Name other separation method Size-exclusion chromatography (non-commercial) Characterization: Protein analysis Protein Yield (µg) per milliliter of starting sample Western Blot Antibody details provided? No Detected EV-associated proteins CD63/ FLOT1/ TSG101 Not detected contaminants Cytochrome C Characterization: RNA analysis RNA analysis Type (RT)(q)PCR/ RNA sequencing/ Capillary electrophoresis (e.g. Bioanalyzer) Database NCBI's Gene Expression Omnibus (GEO) Proteinase treatment No RNAse treatment No Characterization: Lipid analysis No Characterization: Particle analysis NTA Report type Median Reported size (nm) 138.7 EV concentration Yes Particle yield particles per milliliter of starting sample: 4.40E+10 EM EM-type Transmission-EM Image type Close-up

	EV230000	2/2	Bos taurus	Bovine Oviductal Epithelial Cell Organoids	(d)(U)C Filtration Size-exclusion chromatography (non-commercial)	Menjivar NG	2023	67%
Study summary Full title Bovine oviductal organoids: a multi-omics approach to capture the cellular and extracellular molecular response of the oviduct to heat stress. All authors Menjivar NG, Gad A, Thompson RE, Meyers MA, Hollinshead FK, Tesfaye D Journal BMC Genomics Abstract The mammalian oviduct is a complex, fibromuscular organ known for its role in orchestrating a series (show more...)The mammalian oviduct is a complex, fibromuscular organ known for its role in orchestrating a series of timely and dynamic changes to suitably support early embryogenesis. Climate change-induced heat stress (HS) is one of the largest single stressors compromising reproductive function in humans and farm animals via systemic changes in the redox status of the maternal environment, adversely affecting fertilization and early embryonic development. Oviductal organoids represent a unique 3-dimensional, biomimetic model to study the physiology of the oviduct and its subsequent impact on embryo development under various environmental conditions. (hide) EV-METRIC 67% (94th 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 Heat stress (42 degrees celsius) 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 (Differential) (ultra)centrifugation Filtration Size-exclusion chromatography (non-commercial) Protein markers EV: CD63/ FLOT1/ TSG101 non-EV: Cytochrome C Proteomics no Show all info Study aim Identification of content (omics approaches) Sample Species Bos taurus Sample Type Cell culture supernatant EV-producing cells Bovine Oviductal Epithelial Cell Organoids EV-harvesting Medium Serum free medium Separation Method (Differential) (ultra)centrifugation dUC: centrifugation steps Below or equal to 800 g 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: rotor type SW 55 Ti Pelleting: speed (g) 120000 Wash: volume per pellet (ml) 3 Wash: time (min) 70 Wash: Rotor Type SW 55 Ti Wash: speed (g) 120000 Filtration steps 0.2 or 0.22 µm Size-exclusion chromatography Total column volume (mL) 0.1 Sample volume/column (mL) 0.1 Other Name other separation method Size-exclusion chromatography (non-commercial) Characterization: Protein analysis Protein Yield (µg) per milliliter of starting sample Western Blot Antibody details provided? No Detected EV-associated proteins CD63/ FLOT1/ TSG101 Not detected contaminants Cytochrome C Characterization: RNA analysis RNA analysis Type (RT)(q)PCR/ RNA sequencing/ Capillary electrophoresis (e.g. Bioanalyzer) Database NCBI's Gene Expression Omnibus (GEO) Proteinase treatment No RNAse treatment No Characterization: Lipid analysis No Characterization: Particle analysis NTA Report type Median Reported size (nm) 138.4 EV concentration Yes Particle yield particles per milliliter of starting sample: 5.00E+10 EM EM-type Transmission-EM Image type Close-up

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EV-TRACK ID	EV230000
species	Bos taurus
sample type	Cell culture
cell type	Bovine Oviductal Epithelial Cell Organoids
condition	Control condition	Heat stress (42 degrees celsius)
separation protocol	dUC/ Filtration/ Size-exclusion chromatography (non-commercial)	dUC/ Filtration/ Size-exclusion chromatography (non-commercial)
Exp. nr.	1	2
EV-METRIC %	67	67