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Details	EV-TRACK ID	Experiment nr.	Species	Sample type	Separation protocol	First author	Year	EV-METRIC
	EV200129	1/3	Homo sapiens	Blood plasma	(d)(U)C DC Filtration	Maduray, K	2020	33%
Study summary Full title The impact of circulating exosomes derived from early and late onset pre-eclamptic pregnancies on inflammatory cytokine secretion by BeWo cells All authors K Maduray, J Moodley, I Mackraj Journal Eur J Obstet Gynecol Reprod Biol Abstract Objectives: The pathogenesis of pre-eclampsia (PE) is associated with significant maternal and neona (show more...)Objectives: The pathogenesis of pre-eclampsia (PE) is associated with significant maternal and neonatal complications, an increased inflammatory response, placental hypoxia, and endothelial dysfunction, coupled with differential exosomal release profiles with immune modulation effects. Hence, this study evaluated the impact of circulating exosomes derived from early and late-onset pre-eclamptic pregnancies on inflammatory cytokine secretion by BeWo cells. Study design: Exosomes were isolated from plasma obtained from early-onset pre-eclamptic (EOPE; n = 15), late-onset pre-eclamptic (LOPE; n = 15), and gestational age-matched normotensive pregnancies (N ≤ 33 weeks; n = 15 and N ≥ 34 weeks; n = 15). Human BeWo cells were treated with characterized and quantified exosomes (100 μg/mL exosomal protein per pregnant group) for 24 h. The immunoassay method was used to measure the concentration of IL-8, IL-10, leptin, and HIF-α. Results: Exosome administration from women with EOPE and LOPE increased IL-8 and decreased IL-10 expression in BeWo cells. Conclusion: Cumulatively, our data demonstrated that circulating exosomes from the placenta and activated immune cells potentially influence inflammatory cytokine production in pre-eclamptic pregnancies. (hide) EV-METRIC 33% (63rd 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 Blood plasma Sample origin Healthy pregnant Focus vesicles exosome 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 Density cushion Filtration Protein markers EV: CD81/ PLAP/ CD63/ CD9 non-EV: None Proteomics no Show all info Study aim Function Sample Species Homo sapiens Sample Type Blood plasma 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: time(min) 120 Pelleting: rotor type MLA-55 Pelleting: speed (g) 110000 Wash: time (min) 70 Wash: Rotor Type MLA-55 Wash: speed (g) 110000 Filtration steps 0.22µm or 0.2µm Density cushion Density medium Sucrose Characterization: Protein analysis Protein Concentration Method Lowry-based assay Western Blot Detected EV-associated proteins CD9/ CD63/ CD81 ELISA Detected EV-associated proteins CD63/ PLAP Characterization: Lipid analysis No Characterization: Particle analysis NTA Report type Mean Reported size (nm) 103; 104 EV concentration Yes EM EM-type Transmission-EM Image type Wide-field Report size (nm) 30-100

	EV200129	2/3	Homo sapiens	Blood plasma	(d)(U)C DC Filtration	Maduray, K	2020	22%
Study summary Full title The impact of circulating exosomes derived from early and late onset pre-eclamptic pregnancies on inflammatory cytokine secretion by BeWo cells All authors K Maduray, J Moodley, I Mackraj Journal Eur J Obstet Gynecol Reprod Biol Abstract Objectives: The pathogenesis of pre-eclampsia (PE) is associated with significant maternal and neona (show more...)Objectives: The pathogenesis of pre-eclampsia (PE) is associated with significant maternal and neonatal complications, an increased inflammatory response, placental hypoxia, and endothelial dysfunction, coupled with differential exosomal release profiles with immune modulation effects. Hence, this study evaluated the impact of circulating exosomes derived from early and late-onset pre-eclamptic pregnancies on inflammatory cytokine secretion by BeWo cells. Study design: Exosomes were isolated from plasma obtained from early-onset pre-eclamptic (EOPE; n = 15), late-onset pre-eclamptic (LOPE; n = 15), and gestational age-matched normotensive pregnancies (N ≤ 33 weeks; n = 15 and N ≥ 34 weeks; n = 15). Human BeWo cells were treated with characterized and quantified exosomes (100 μg/mL exosomal protein per pregnant group) for 24 h. The immunoassay method was used to measure the concentration of IL-8, IL-10, leptin, and HIF-α. Results: Exosome administration from women with EOPE and LOPE increased IL-8 and decreased IL-10 expression in BeWo cells. Conclusion: Cumulatively, our data demonstrated that circulating exosomes from the placenta and activated immune cells potentially influence inflammatory cytokine production in pre-eclamptic pregnancies. (hide) EV-METRIC 22% (49th 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 Blood plasma Sample origin Early onset pre-eclampsia Focus vesicles exosome 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 Density cushion Filtration Protein markers EV: CD81/ PLAP/ CD63/ CD9 non-EV: None Proteomics no Show all info Study aim Function Sample Species Homo sapiens Sample Type Blood plasma 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: time(min) 120 Pelleting: rotor type MLA-55 Pelleting: speed (g) 110000 Wash: time (min) 70 Wash: Rotor Type MLA-55 Wash: speed (g) 110000 Filtration steps 0.22µm or 0.2µm Density cushion Density medium Sucrose Characterization: Protein analysis Protein Concentration Method Lowry-based assay Western Blot Detected EV-associated proteins CD9/ CD63/ CD81 ELISA Detected EV-associated proteins CD63/ PLAP Characterization: Lipid analysis No Characterization: Particle analysis NTA Report type Mean Reported size (nm) 101 EV concentration Yes

	EV200129	3/3	Homo sapiens	Blood plasma	(d)(U)C DC Filtration	Maduray, K	2020	22%
Study summary Full title The impact of circulating exosomes derived from early and late onset pre-eclamptic pregnancies on inflammatory cytokine secretion by BeWo cells All authors K Maduray, J Moodley, I Mackraj Journal Eur J Obstet Gynecol Reprod Biol Abstract Objectives: The pathogenesis of pre-eclampsia (PE) is associated with significant maternal and neona (show more...)Objectives: The pathogenesis of pre-eclampsia (PE) is associated with significant maternal and neonatal complications, an increased inflammatory response, placental hypoxia, and endothelial dysfunction, coupled with differential exosomal release profiles with immune modulation effects. Hence, this study evaluated the impact of circulating exosomes derived from early and late-onset pre-eclamptic pregnancies on inflammatory cytokine secretion by BeWo cells. Study design: Exosomes were isolated from plasma obtained from early-onset pre-eclamptic (EOPE; n = 15), late-onset pre-eclamptic (LOPE; n = 15), and gestational age-matched normotensive pregnancies (N ≤ 33 weeks; n = 15 and N ≥ 34 weeks; n = 15). Human BeWo cells were treated with characterized and quantified exosomes (100 μg/mL exosomal protein per pregnant group) for 24 h. The immunoassay method was used to measure the concentration of IL-8, IL-10, leptin, and HIF-α. Results: Exosome administration from women with EOPE and LOPE increased IL-8 and decreased IL-10 expression in BeWo cells. Conclusion: Cumulatively, our data demonstrated that circulating exosomes from the placenta and activated immune cells potentially influence inflammatory cytokine production in pre-eclamptic pregnancies. (hide) EV-METRIC 22% (49th 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 Blood plasma Sample origin Late onset pre-eclampsia Focus vesicles exosome 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 Density cushion Filtration Protein markers EV: CD81/ PLAP/ CD63/ CD9 non-EV: None Proteomics no Show all info Study aim Function Sample Species Homo sapiens Sample Type Blood plasma 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: time(min) 120 Pelleting: rotor type MLA-55 Pelleting: speed (g) 110000 Wash: time (min) 70 Wash: Rotor Type MLA-55 Wash: speed (g) 110000 Filtration steps 0.22µm or 0.2µm Density cushion Density medium Sucrose Characterization: Protein analysis Protein Concentration Method Lowry-based assay Western Blot Detected EV-associated proteins CD9/ CD63/ CD81 ELISA Detected EV-associated proteins CD63/ PLAP Characterization: Lipid analysis No Characterization: Particle analysis NTA Report type Mean Reported size (nm) 102 EV concentration Yes

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EV-TRACK ID	EV200129
species	Homo sapiens
sample type	Blood plasma
condition	Healthy pregnant	Early onset pre-eclampsia	Late onset pre-eclampsia
separation protocol	dUC DC Filtration	dUC DC Filtration	dUC DC Filtration
Exp. nr.	1	2	3
EV-METRIC %	33	22	22