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Details	EV-TRACK ID	Experiment nr.	Species	Sample type	Separation protocol	First author	Year	EV-METRIC
	EV200036	1/16	Homo sapiens	human skin primary fibroblasts	DG (d)(U)C qEV	Juan Antonio Fafián-Labora	2020	78%
Study summary Full title Small Extracellular Vesicles Have GST Activity and Ameliorate Senescence-Related Tissue Damage All authors Juan Antonio Fafián-Labora, Jose Antonio Rodríguez-Navarro, Ana O'Loghlen Journal Cell metab Abstract Aging is a process of cellular and tissue dysfunction characterized by different hallmarks, includin (show more...)Aging is a process of cellular and tissue dysfunction characterized by different hallmarks, including cellular senescence. However, there is proof that certain features of aging and senescence can be ameliorated. Here, we provide evidence that small extracellular vesicles (sEVs) isolated from primary fibroblasts of young human donors ameliorate certain biomarkers of senescence in cells derived from old and Hutchinson-Gilford progeria syndrome donors. Importantly, sEVs from young cells ameliorate senescence in a variety of tissues in old mice. Mechanistically, we identified sEVs to have intrinsic glutathione-S-transferase activity partially due to the high levels of expression of the glutathione-related protein (GSTM2). Transfection of recombinant GSTM2 into sEVs derived from old fibroblasts restores their antioxidant capacity. sEVs increase the levels of reduced glutathione and decrease oxidative stress and lipid peroxidation both in vivo and in vitro. Altogether, our data provide an indication of the potential of sEVs as regenerative therapy in aging. (hide) EV-METRIC 78% (97th 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 Young donors 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 Commercial method Protein markers EV: Alix/ TSG101/ GSTM2 non-EV: Calnexin/ Actin-beta Proteomics no EV density (g/ml) 1.074-1.106 Show all info Study aim Function Sample Species Homo sapiens Sample Type Cell culture supernatant EV-producing cells human skin primary fibroblasts EV-harvesting Medium EV-depleted medium Preparation of EDS overnight (16h) at >= 100,000g 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) 80 Pelleting: rotor type T-865 Pelleting: speed (g) 100000 Wash: volume per pellet (ml) 15 Wash: time (min) 80 Wash: Rotor Type T-865 Wash: speed (g) 100000 Density gradient Type Discontinuous Number of initial discontinuous layers 3 Lowest density fraction 10% Highest density fraction 60% Total gradient volume, incl. sample (mL) 5.5 Sample volume (mL) 1.5 Orientation Bottom-up Rotor type T-865 Speed (g) 100000 Duration (min) 720 Fraction volume (mL) 0.7 Fraction processing Centrifugation Pelleting: volume per fraction 15 Pelleting: duration (min) 80 Pelleting: rotor type T-865 Pelleting: speed (g) 100000 Pelleting-wash: volume per pellet (mL) 15 Pelleting-wash: duration (min) 80 Pelleting-wash: speed (g) T-865 Commercial kit qEV EV-subtype Distinction between multiple subtypes Size Used subtypes <200 nm Characterization: Protein analysis Protein Concentration Method Not determined Western Blot Detected EV-associated proteins Alix/ TSG101/ GSTM2 Not detected contaminants Calnexin/ Actin-beta Characterization: Lipid analysis No Characterization: Particle analysis NTA Report type Mean Reported size (nm) <200 EV concentration Yes Particle yield Number of particles of starting sample E08-E09

	EV200036	3/16	Homo sapiens	human skin primary fibroblasts	DG (d)(U)C qEV	Juan Antonio Fafián-Labora	2020	78%
Study summary Full title Small Extracellular Vesicles Have GST Activity and Ameliorate Senescence-Related Tissue Damage All authors Juan Antonio Fafián-Labora, Jose Antonio Rodríguez-Navarro, Ana O'Loghlen Journal Cell metab Abstract Aging is a process of cellular and tissue dysfunction characterized by different hallmarks, includin (show more...)Aging is a process of cellular and tissue dysfunction characterized by different hallmarks, including cellular senescence. However, there is proof that certain features of aging and senescence can be ameliorated. Here, we provide evidence that small extracellular vesicles (sEVs) isolated from primary fibroblasts of young human donors ameliorate certain biomarkers of senescence in cells derived from old and Hutchinson-Gilford progeria syndrome donors. Importantly, sEVs from young cells ameliorate senescence in a variety of tissues in old mice. Mechanistically, we identified sEVs to have intrinsic glutathione-S-transferase activity partially due to the high levels of expression of the glutathione-related protein (GSTM2). Transfection of recombinant GSTM2 into sEVs derived from old fibroblasts restores their antioxidant capacity. sEVs increase the levels of reduced glutathione and decrease oxidative stress and lipid peroxidation both in vivo and in vitro. Altogether, our data provide an indication of the potential of sEVs as regenerative therapy in aging. (hide) EV-METRIC 78% (97th 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 Old donors 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 Commercial method Protein markers EV: Alix/ TSG101/ GSTM2 non-EV: Calnexin/ Actin-beta Proteomics no EV density (g/ml) 1.074-1.106 Show all info Study aim Function Sample Species Homo sapiens Sample Type Cell culture supernatant EV-producing cells human skin primary fibroblasts EV-harvesting Medium EV-depleted medium Preparation of EDS overnight (16h) at >= 100,000g 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) 80 Pelleting: rotor type T-865 Pelleting: speed (g) 100000 Wash: volume per pellet (ml) 15 Wash: time (min) 80 Wash: Rotor Type T-865 Wash: speed (g) 100000 Density gradient Type Discontinuous Number of initial discontinuous layers 3 Lowest density fraction 10% Highest density fraction 60% Total gradient volume, incl. sample (mL) 5.5 Sample volume (mL) 1.5 Orientation Bottom-up Rotor type T-865 Speed (g) 100000 Duration (min) 720 Fraction volume (mL) 0.7 Fraction processing Centrifugation Pelleting: volume per fraction 15 Pelleting: duration (min) 80 Pelleting: rotor type T-865 Pelleting: speed (g) 100000 Pelleting-wash: volume per pellet (mL) 15 Pelleting-wash: duration (min) 80 Pelleting-wash: speed (g) T-865 Commercial kit qEV EV-subtype Distinction between multiple subtypes Size Used subtypes <200 nm Characterization: Protein analysis Protein Concentration Method Not determined Western Blot Detected EV-associated proteins Alix/ TSG101/ GSTM2 Not detected contaminants Calnexin/ Actin-beta Characterization: Lipid analysis No Characterization: Particle analysis NTA Report type Mean Reported size (nm) <200 EV concentration Yes Particle yield Number of particles of starting sample E08-E09

	EV200036	5/16	Homo sapiens	human skin primary fibroblasts	DG (d)(U)C qEV	Juan Antonio Fafián-Labora	2020	56%
Study summary Full title Small Extracellular Vesicles Have GST Activity and Ameliorate Senescence-Related Tissue Damage All authors Juan Antonio Fafián-Labora, Jose Antonio Rodríguez-Navarro, Ana O'Loghlen Journal Cell metab Abstract Aging is a process of cellular and tissue dysfunction characterized by different hallmarks, includin (show more...)Aging is a process of cellular and tissue dysfunction characterized by different hallmarks, including cellular senescence. However, there is proof that certain features of aging and senescence can be ameliorated. Here, we provide evidence that small extracellular vesicles (sEVs) isolated from primary fibroblasts of young human donors ameliorate certain biomarkers of senescence in cells derived from old and Hutchinson-Gilford progeria syndrome donors. Importantly, sEVs from young cells ameliorate senescence in a variety of tissues in old mice. Mechanistically, we identified sEVs to have intrinsic glutathione-S-transferase activity partially due to the high levels of expression of the glutathione-related protein (GSTM2). Transfection of recombinant GSTM2 into sEVs derived from old fibroblasts restores their antioxidant capacity. sEVs increase the levels of reduced glutathione and decrease oxidative stress and lipid peroxidation both in vivo and in vitro. Altogether, our data provide an indication of the potential of sEVs as regenerative therapy in aging. (hide) EV-METRIC 56% (90th 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 Progeria patients 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 Commercial method Protein markers EV: Alix/ GSTM2 non-EV: None Proteomics no EV density (g/ml) 1.074-1.106 Show all info Study aim Function Sample Species Homo sapiens Sample Type Cell culture supernatant EV-producing cells human skin primary fibroblasts EV-harvesting Medium EV-depleted medium Preparation of EDS overnight (16h) at >= 100,000g 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) 80 Pelleting: rotor type T-865 Pelleting: speed (g) 100000 Wash: volume per pellet (ml) 15 Wash: time (min) 80 Wash: Rotor Type T-865 Wash: speed (g) 100000 Density gradient Type Discontinuous Number of initial discontinuous layers 3 Lowest density fraction 10% Highest density fraction 60% Total gradient volume, incl. sample (mL) 5.5 Sample volume (mL) 1.5 Orientation Bottom-up Rotor type T-865 Speed (g) 100000 Duration (min) 720 Fraction volume (mL) 0.7 Fraction processing Centrifugation Pelleting: volume per fraction 15 Pelleting: duration (min) 80 Pelleting: rotor type T-865 Pelleting: speed (g) 100000 Pelleting-wash: volume per pellet (mL) 15 Pelleting-wash: duration (min) 80 Pelleting-wash: speed (g) T-865 Commercial kit qEV EV-subtype Distinction between multiple subtypes Size Used subtypes <200 nm Characterization: Protein analysis Protein Concentration Method Not determined Western Blot Detected EV-associated proteins Alix/ GSTM2 Characterization: Lipid analysis No Characterization: Particle analysis NTA Report type Mean Reported size (nm) <200 EV concentration Yes Particle yield Number of particles of starting sample E08-E09

	EV200036	7/16	Homo sapiens	primary human foreskin fibroblasts	DG (d)(U)C qEV	Juan Antonio Fafián-Labora	2020	56%
Study summary Full title Small Extracellular Vesicles Have GST Activity and Ameliorate Senescence-Related Tissue Damage All authors Juan Antonio Fafián-Labora, Jose Antonio Rodríguez-Navarro, Ana O'Loghlen Journal Cell metab Abstract Aging is a process of cellular and tissue dysfunction characterized by different hallmarks, includin (show more...)Aging is a process of cellular and tissue dysfunction characterized by different hallmarks, including cellular senescence. However, there is proof that certain features of aging and senescence can be ameliorated. Here, we provide evidence that small extracellular vesicles (sEVs) isolated from primary fibroblasts of young human donors ameliorate certain biomarkers of senescence in cells derived from old and Hutchinson-Gilford progeria syndrome donors. Importantly, sEVs from young cells ameliorate senescence in a variety of tissues in old mice. Mechanistically, we identified sEVs to have intrinsic glutathione-S-transferase activity partially due to the high levels of expression of the glutathione-related protein (GSTM2). Transfection of recombinant GSTM2 into sEVs derived from old fibroblasts restores their antioxidant capacity. sEVs increase the levels of reduced glutathione and decrease oxidative stress and lipid peroxidation both in vivo and in vitro. Altogether, our data provide an indication of the potential of sEVs as regenerative therapy in aging. (hide) EV-METRIC 56% (90th 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 Commercial method Protein markers EV: Alix/ GSTM2 non-EV: None Proteomics no EV density (g/ml) 1.074-1.106 Show all info Study aim Function Sample Species Homo sapiens Sample Type Cell culture supernatant EV-producing cells primary human foreskin fibroblasts EV-harvesting Medium EV-depleted medium Preparation of EDS overnight (16h) at >= 100,000g 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) 80 Pelleting: rotor type T-865 Pelleting: speed (g) 100000 Wash: volume per pellet (ml) 15 Wash: time (min) 80 Wash: Rotor Type T-865 Wash: speed (g) 100000 Density gradient Type Discontinuous Number of initial discontinuous layers 3 Lowest density fraction 10% Highest density fraction 60% Total gradient volume, incl. sample (mL) 5.5 Sample volume (mL) 1.5 Orientation Bottom-up Rotor type T-865 Speed (g) 100000 Duration (min) 720 Fraction volume (mL) 0.7 Fraction processing Centrifugation Pelleting: volume per fraction 15 Pelleting: duration (min) 80 Pelleting: rotor type T-865 Pelleting: speed (g) 100000 Pelleting-wash: volume per pellet (mL) 15 Pelleting-wash: duration (min) 80 Pelleting-wash: speed (g) T-865 Commercial kit qEV EV-subtype Distinction between multiple subtypes Size Used subtypes <200 nm Characterization: Protein analysis Protein Concentration Method Not determined Western Blot Detected EV-associated proteins Alix/ GSTM2 Characterization: Lipid analysis No Characterization: Particle analysis NTA Report type Mean Reported size (nm) <200 EV concentration Yes Particle yield Number of particles of starting sample E08-E09

	EV200036	9/16	Homo sapiens	primary human foreskin fibroblasts	DG (d)(U)C qEV	Juan Antonio Fafián-Labora	2020	56%
Study summary Full title Small Extracellular Vesicles Have GST Activity and Ameliorate Senescence-Related Tissue Damage All authors Juan Antonio Fafián-Labora, Jose Antonio Rodríguez-Navarro, Ana O'Loghlen Journal Cell metab Abstract Aging is a process of cellular and tissue dysfunction characterized by different hallmarks, includin (show more...)Aging is a process of cellular and tissue dysfunction characterized by different hallmarks, including cellular senescence. However, there is proof that certain features of aging and senescence can be ameliorated. Here, we provide evidence that small extracellular vesicles (sEVs) isolated from primary fibroblasts of young human donors ameliorate certain biomarkers of senescence in cells derived from old and Hutchinson-Gilford progeria syndrome donors. Importantly, sEVs from young cells ameliorate senescence in a variety of tissues in old mice. Mechanistically, we identified sEVs to have intrinsic glutathione-S-transferase activity partially due to the high levels of expression of the glutathione-related protein (GSTM2). Transfection of recombinant GSTM2 into sEVs derived from old fibroblasts restores their antioxidant capacity. sEVs increase the levels of reduced glutathione and decrease oxidative stress and lipid peroxidation both in vivo and in vitro. Altogether, our data provide an indication of the potential of sEVs as regenerative therapy in aging. (hide) EV-METRIC 56% (90th 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 iRas 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 Commercial method Protein markers EV: Alix/ GSTM2 non-EV: None Proteomics no EV density (g/ml) 1.074-1.106 Show all info Study aim Function Sample Species Homo sapiens Sample Type Cell culture supernatant EV-producing cells primary human foreskin fibroblasts EV-harvesting Medium EV-depleted medium Preparation of EDS overnight (16h) at >= 100,000g 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) 80 Pelleting: rotor type T-865 Pelleting: speed (g) 100000 Wash: volume per pellet (ml) 15 Wash: time (min) 80 Wash: Rotor Type T-865 Wash: speed (g) 100000 Density gradient Type Discontinuous Number of initial discontinuous layers 3 Lowest density fraction 10% Highest density fraction 60% Total gradient volume, incl. sample (mL) 5.5 Sample volume (mL) 1.5 Orientation Bottom-up Rotor type T-865 Speed (g) 100000 Duration (min) 720 Fraction volume (mL) 0.7 Fraction processing Centrifugation Pelleting: volume per fraction 15 Pelleting: duration (min) 80 Pelleting: rotor type T-865 Pelleting: speed (g) 100000 Pelleting-wash: volume per pellet (mL) 15 Pelleting-wash: duration (min) 80 Pelleting-wash: speed (g) T-865 Commercial kit qEV EV-subtype Distinction between multiple subtypes Size Used subtypes <200 nm Characterization: Protein analysis Protein Concentration Method Not determined Western Blot Detected EV-associated proteins Alix/ GSTM2 Characterization: Lipid analysis No Characterization: Particle analysis NTA Report type Mean Reported size (nm) <200 EV concentration Yes Particle yield Number of particles of starting sample E08-E09

	EV200036	11/16	Homo sapiens	primary human foreskin fibroblasts	DG (d)(U)C qEV	Juan Antonio Fafián-Labora	2020	56%
Study summary Full title Small Extracellular Vesicles Have GST Activity and Ameliorate Senescence-Related Tissue Damage All authors Juan Antonio Fafián-Labora, Jose Antonio Rodríguez-Navarro, Ana O'Loghlen Journal Cell metab Abstract Aging is a process of cellular and tissue dysfunction characterized by different hallmarks, includin (show more...)Aging is a process of cellular and tissue dysfunction characterized by different hallmarks, including cellular senescence. However, there is proof that certain features of aging and senescence can be ameliorated. Here, we provide evidence that small extracellular vesicles (sEVs) isolated from primary fibroblasts of young human donors ameliorate certain biomarkers of senescence in cells derived from old and Hutchinson-Gilford progeria syndrome donors. Importantly, sEVs from young cells ameliorate senescence in a variety of tissues in old mice. Mechanistically, we identified sEVs to have intrinsic glutathione-S-transferase activity partially due to the high levels of expression of the glutathione-related protein (GSTM2). Transfection of recombinant GSTM2 into sEVs derived from old fibroblasts restores their antioxidant capacity. sEVs increase the levels of reduced glutathione and decrease oxidative stress and lipid peroxidation both in vivo and in vitro. Altogether, our data provide an indication of the potential of sEVs as regenerative therapy in aging. (hide) EV-METRIC 56% (90th 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 iRas+GSTM2 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 Commercial method Protein markers EV: Alix/ GSTM2 non-EV: None Proteomics no EV density (g/ml) 1.074-1.106 Show all info Study aim Function Sample Species Homo sapiens Sample Type Cell culture supernatant EV-producing cells primary human foreskin fibroblasts EV-harvesting Medium EV-depleted medium Preparation of EDS overnight (16h) at >= 100,000g 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) 80 Pelleting: rotor type T-865 Pelleting: speed (g) 100000 Wash: volume per pellet (ml) 15 Wash: time (min) 80 Wash: Rotor Type T-865 Wash: speed (g) 100000 Density gradient Type Discontinuous Number of initial discontinuous layers 3 Lowest density fraction 10% Highest density fraction 60% Total gradient volume, incl. sample (mL) 5.5 Sample volume (mL) 1.5 Orientation Bottom-up Rotor type T-865 Speed (g) 100000 Duration (min) 720 Fraction volume (mL) 0.7 Fraction processing Centrifugation Pelleting: volume per fraction 15 Pelleting: duration (min) 80 Pelleting: rotor type T-865 Pelleting: speed (g) 100000 Pelleting-wash: volume per pellet (mL) 15 Pelleting-wash: duration (min) 80 Pelleting-wash: speed (g) T-865 Commercial kit qEV EV-subtype Distinction between multiple subtypes Size Used subtypes <200 nm Characterization: Protein analysis Protein Concentration Method Not determined Western Blot Detected EV-associated proteins Alix/ GSTM2 Characterization: Lipid analysis No Characterization: Particle analysis NTA Report type Mean Reported size (nm) <200 EV concentration Yes Particle yield Number of particles of starting sample E08-E09

	EV200036	13/16	Homo sapiens	primary human foreskin fibroblasts	DG (d)(U)C qEV	Juan Antonio Fafián-Labora	2020	56%
Study summary Full title Small Extracellular Vesicles Have GST Activity and Ameliorate Senescence-Related Tissue Damage All authors Juan Antonio Fafián-Labora, Jose Antonio Rodríguez-Navarro, Ana O'Loghlen Journal Cell metab Abstract Aging is a process of cellular and tissue dysfunction characterized by different hallmarks, includin (show more...)Aging is a process of cellular and tissue dysfunction characterized by different hallmarks, including cellular senescence. However, there is proof that certain features of aging and senescence can be ameliorated. Here, we provide evidence that small extracellular vesicles (sEVs) isolated from primary fibroblasts of young human donors ameliorate certain biomarkers of senescence in cells derived from old and Hutchinson-Gilford progeria syndrome donors. Importantly, sEVs from young cells ameliorate senescence in a variety of tissues in old mice. Mechanistically, we identified sEVs to have intrinsic glutathione-S-transferase activity partially due to the high levels of expression of the glutathione-related protein (GSTM2). Transfection of recombinant GSTM2 into sEVs derived from old fibroblasts restores their antioxidant capacity. sEVs increase the levels of reduced glutathione and decrease oxidative stress and lipid peroxidation both in vivo and in vitro. Altogether, our data provide an indication of the potential of sEVs as regenerative therapy in aging. (hide) EV-METRIC 56% (90th 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 iC+GSTM2 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 Commercial method Protein markers EV: Alix/ GSTM2 non-EV: None Proteomics no EV density (g/ml) 1.074-1.106 Show all info Study aim Function Sample Species Homo sapiens Sample Type Cell culture supernatant EV-producing cells primary human foreskin fibroblasts EV-harvesting Medium EV-depleted medium Preparation of EDS overnight (16h) at >= 100,000g 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) 80 Pelleting: rotor type T-865 Pelleting: speed (g) 100000 Wash: volume per pellet (ml) 15 Wash: time (min) 80 Wash: Rotor Type T-865 Wash: speed (g) 100000 Density gradient Type Discontinuous Number of initial discontinuous layers 3 Lowest density fraction 10% Highest density fraction 60% Total gradient volume, incl. sample (mL) 5.5 Sample volume (mL) 1.5 Orientation Bottom-up Rotor type T-865 Speed (g) 100000 Duration (min) 720 Fraction volume (mL) 0.7 Fraction processing Centrifugation Pelleting: volume per fraction 15 Pelleting: duration (min) 80 Pelleting: rotor type T-865 Pelleting: speed (g) 100000 Pelleting-wash: volume per pellet (mL) 15 Pelleting-wash: duration (min) 80 Pelleting-wash: speed (g) T-865 Commercial kit qEV EV-subtype Distinction between multiple subtypes Size Used subtypes <200 nm Characterization: Protein analysis Protein Concentration Method Not determined Western Blot Detected EV-associated proteins Alix/ GSTM2 Characterization: Lipid analysis No Characterization: Particle analysis NTA Report type Mean Reported size (nm) <200 EV concentration Yes Particle yield Number of particles of starting sample E08-E09

	EV200036	15/16	Homo sapiens	primary human foreskin fibroblasts	DG (d)(U)C qEV	Juan Antonio Fafián-Labora	2020	56%
Study summary Full title Small Extracellular Vesicles Have GST Activity and Ameliorate Senescence-Related Tissue Damage All authors Juan Antonio Fafián-Labora, Jose Antonio Rodríguez-Navarro, Ana O'Loghlen Journal Cell metab Abstract Aging is a process of cellular and tissue dysfunction characterized by different hallmarks, includin (show more...)Aging is a process of cellular and tissue dysfunction characterized by different hallmarks, including cellular senescence. However, there is proof that certain features of aging and senescence can be ameliorated. Here, we provide evidence that small extracellular vesicles (sEVs) isolated from primary fibroblasts of young human donors ameliorate certain biomarkers of senescence in cells derived from old and Hutchinson-Gilford progeria syndrome donors. Importantly, sEVs from young cells ameliorate senescence in a variety of tissues in old mice. Mechanistically, we identified sEVs to have intrinsic glutathione-S-transferase activity partially due to the high levels of expression of the glutathione-related protein (GSTM2). Transfection of recombinant GSTM2 into sEVs derived from old fibroblasts restores their antioxidant capacity. sEVs increase the levels of reduced glutathione and decrease oxidative stress and lipid peroxidation both in vivo and in vitro. Altogether, our data provide an indication of the potential of sEVs as regenerative therapy in aging. (hide) EV-METRIC 56% (90th 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 mCherry-CD63 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 Commercial method Protein markers EV: Alix/ GSTM2 non-EV: None Proteomics no EV density (g/ml) 1.074-1.106 Show all info Study aim Function Sample Species Homo sapiens Sample Type Cell culture supernatant EV-producing cells primary human foreskin fibroblasts EV-harvesting Medium EV-depleted medium Preparation of EDS overnight (16h) at >= 100,000g 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) 80 Pelleting: rotor type T-865 Pelleting: speed (g) 100000 Wash: volume per pellet (ml) 15 Wash: time (min) 80 Wash: Rotor Type T-865 Wash: speed (g) 100000 Density gradient Type Discontinuous Number of initial discontinuous layers 3 Lowest density fraction 10% Highest density fraction 60% Total gradient volume, incl. sample (mL) 5.5 Sample volume (mL) 1.5 Orientation Bottom-up Rotor type T-865 Speed (g) 100000 Duration (min) 720 Fraction volume (mL) 0.7 Fraction processing Centrifugation Pelleting: volume per fraction 15 Pelleting: duration (min) 80 Pelleting: rotor type T-865 Pelleting: speed (g) 100000 Pelleting-wash: volume per pellet (mL) 15 Pelleting-wash: duration (min) 80 Pelleting-wash: speed (g) T-865 Commercial kit qEV EV-subtype Distinction between multiple subtypes Size Used subtypes <200 nm Characterization: Protein analysis Protein Concentration Method Not determined Western Blot Detected EV-associated proteins Alix/ GSTM2 Characterization: Lipid analysis No Characterization: Particle analysis NTA Report type Mean Reported size (nm) <200 EV concentration Yes Particle yield Number of particles of starting sample E08-E09

	EV200036	2/16	Homo sapiens	human skin primary fibroblasts	DG (d)(U)C qEV	Juan Antonio Fafián-Labora	2020	43%
Study summary Full title Small Extracellular Vesicles Have GST Activity and Ameliorate Senescence-Related Tissue Damage All authors Juan Antonio Fafián-Labora, Jose Antonio Rodríguez-Navarro, Ana O'Loghlen Journal Cell metab Abstract Aging is a process of cellular and tissue dysfunction characterized by different hallmarks, includin (show more...)Aging is a process of cellular and tissue dysfunction characterized by different hallmarks, including cellular senescence. However, there is proof that certain features of aging and senescence can be ameliorated. Here, we provide evidence that small extracellular vesicles (sEVs) isolated from primary fibroblasts of young human donors ameliorate certain biomarkers of senescence in cells derived from old and Hutchinson-Gilford progeria syndrome donors. Importantly, sEVs from young cells ameliorate senescence in a variety of tissues in old mice. Mechanistically, we identified sEVs to have intrinsic glutathione-S-transferase activity partially due to the high levels of expression of the glutathione-related protein (GSTM2). Transfection of recombinant GSTM2 into sEVs derived from old fibroblasts restores their antioxidant capacity. sEVs increase the levels of reduced glutathione and decrease oxidative stress and lipid peroxidation both in vivo and in vitro. Altogether, our data provide an indication of the potential of sEVs as regenerative therapy in aging. (hide) EV-METRIC 43% (81st 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 Young donors 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 Commercial method Protein markers EV: None non-EV: None Proteomics no EV density (g/ml) 1.074-1.106 Show all info Study aim Function Sample Species Homo sapiens Sample Type Cell culture supernatant EV-producing cells human skin primary fibroblasts EV-harvesting Medium EV-depleted medium Preparation of EDS overnight (16h) at >= 100,000g 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) 80 Pelleting: rotor type T-865 Pelleting: speed (g) 100000 Wash: volume per pellet (ml) 15 Wash: time (min) 80 Wash: Rotor Type T-865 Wash: speed (g) 100000 Density gradient Type Discontinuous Number of initial discontinuous layers 3 Lowest density fraction 10% Highest density fraction 60% Total gradient volume, incl. sample (mL) 5.5 Sample volume (mL) 1.5 Orientation Bottom-up Rotor type T-865 Speed (g) 100000 Duration (min) 720 Fraction volume (mL) 0.7 Fraction processing Centrifugation Pelleting: volume per fraction 15 Pelleting: duration (min) 80 Pelleting: rotor type T-865 Pelleting: speed (g) 100000 Pelleting-wash: volume per pellet (mL) 15 Pelleting-wash: duration (min) 80 Pelleting-wash: speed (g) T-865 Commercial kit qEV EV-subtype Distinction between multiple subtypes Size Used subtypes >200 nm Characterization: Protein analysis None Protein Concentration Method Not determined Characterization: Lipid analysis No Characterization: Particle analysis NTA Report type Mean Reported size (nm) >200 EV concentration Yes Particle yield Number of particles of starting sample E08-E09

	EV200036	4/16	Homo sapiens	human skin primary fibroblasts	DG (d)(U)C qEV	Juan Antonio Fafián-Labora	2020	43%
Study summary Full title Small Extracellular Vesicles Have GST Activity and Ameliorate Senescence-Related Tissue Damage All authors Juan Antonio Fafián-Labora, Jose Antonio Rodríguez-Navarro, Ana O'Loghlen Journal Cell metab Abstract Aging is a process of cellular and tissue dysfunction characterized by different hallmarks, includin (show more...)Aging is a process of cellular and tissue dysfunction characterized by different hallmarks, including cellular senescence. However, there is proof that certain features of aging and senescence can be ameliorated. Here, we provide evidence that small extracellular vesicles (sEVs) isolated from primary fibroblasts of young human donors ameliorate certain biomarkers of senescence in cells derived from old and Hutchinson-Gilford progeria syndrome donors. Importantly, sEVs from young cells ameliorate senescence in a variety of tissues in old mice. Mechanistically, we identified sEVs to have intrinsic glutathione-S-transferase activity partially due to the high levels of expression of the glutathione-related protein (GSTM2). Transfection of recombinant GSTM2 into sEVs derived from old fibroblasts restores their antioxidant capacity. sEVs increase the levels of reduced glutathione and decrease oxidative stress and lipid peroxidation both in vivo and in vitro. Altogether, our data provide an indication of the potential of sEVs as regenerative therapy in aging. (hide) EV-METRIC 43% (81st 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 Old donors 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 Commercial method Protein markers EV: None non-EV: None Proteomics no EV density (g/ml) 1.074-1.106 Show all info Study aim Function Sample Species Homo sapiens Sample Type Cell culture supernatant EV-producing cells human skin primary fibroblasts EV-harvesting Medium EV-depleted medium Preparation of EDS overnight (16h) at >= 100,000g 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) 80 Pelleting: rotor type T-865 Pelleting: speed (g) 100000 Wash: volume per pellet (ml) 15 Wash: time (min) 80 Wash: Rotor Type T-865 Wash: speed (g) 100000 Density gradient Type Discontinuous Number of initial discontinuous layers 3 Lowest density fraction 10% Highest density fraction 60% Total gradient volume, incl. sample (mL) 5.5 Sample volume (mL) 1.5 Orientation Bottom-up Rotor type T-865 Speed (g) 100000 Duration (min) 720 Fraction volume (mL) 0.7 Fraction processing Centrifugation Pelleting: volume per fraction 15 Pelleting: duration (min) 80 Pelleting: rotor type T-865 Pelleting: speed (g) 100000 Pelleting-wash: volume per pellet (mL) 15 Pelleting-wash: duration (min) 80 Pelleting-wash: speed (g) T-865 Commercial kit qEV EV-subtype Distinction between multiple subtypes Size Used subtypes >200 nm Characterization: Protein analysis None Protein Concentration Method Not determined Characterization: Lipid analysis No Characterization: Particle analysis NTA Report type Mean Reported size (nm) >200 EV concentration Yes Particle yield Number of particles of starting sample E08-E09

	EV200036	6/16	Homo sapiens	human skin primary fibroblasts	DG (d)(U)C qEV	Juan Antonio Fafián-Labora	2020	43%
Study summary Full title Small Extracellular Vesicles Have GST Activity and Ameliorate Senescence-Related Tissue Damage All authors Juan Antonio Fafián-Labora, Jose Antonio Rodríguez-Navarro, Ana O'Loghlen Journal Cell metab Abstract Aging is a process of cellular and tissue dysfunction characterized by different hallmarks, includin (show more...)Aging is a process of cellular and tissue dysfunction characterized by different hallmarks, including cellular senescence. However, there is proof that certain features of aging and senescence can be ameliorated. Here, we provide evidence that small extracellular vesicles (sEVs) isolated from primary fibroblasts of young human donors ameliorate certain biomarkers of senescence in cells derived from old and Hutchinson-Gilford progeria syndrome donors. Importantly, sEVs from young cells ameliorate senescence in a variety of tissues in old mice. Mechanistically, we identified sEVs to have intrinsic glutathione-S-transferase activity partially due to the high levels of expression of the glutathione-related protein (GSTM2). Transfection of recombinant GSTM2 into sEVs derived from old fibroblasts restores their antioxidant capacity. sEVs increase the levels of reduced glutathione and decrease oxidative stress and lipid peroxidation both in vivo and in vitro. Altogether, our data provide an indication of the potential of sEVs as regenerative therapy in aging. (hide) EV-METRIC 43% (81st 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 Progeria patients 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 Commercial method Protein markers EV: None non-EV: None Proteomics no EV density (g/ml) 1.074-1.106 Show all info Study aim Function Sample Species Homo sapiens Sample Type Cell culture supernatant EV-producing cells human skin primary fibroblasts EV-harvesting Medium EV-depleted medium Preparation of EDS overnight (16h) at >= 100,000g 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) 80 Pelleting: rotor type T-865 Pelleting: speed (g) 100000 Wash: volume per pellet (ml) 15 Wash: time (min) 80 Wash: Rotor Type T-865 Wash: speed (g) 100000 Density gradient Type Discontinuous Number of initial discontinuous layers 3 Lowest density fraction 10% Highest density fraction 60% Total gradient volume, incl. sample (mL) 5.5 Sample volume (mL) 1.5 Orientation Bottom-up Rotor type T-865 Speed (g) 100000 Duration (min) 720 Fraction volume (mL) 0.7 Fraction processing Centrifugation Pelleting: volume per fraction 15 Pelleting: duration (min) 80 Pelleting: rotor type T-865 Pelleting: speed (g) 100000 Pelleting-wash: volume per pellet (mL) 15 Pelleting-wash: duration (min) 80 Pelleting-wash: speed (g) T-865 Commercial kit qEV EV-subtype Distinction between multiple subtypes Size Used subtypes >200 nm Characterization: Protein analysis None Protein Concentration Method Not determined Characterization: Lipid analysis No Characterization: Particle analysis None

	EV200036	8/16	Homo sapiens	primary human foreskin fibroblasts	DG (d)(U)C qEV	Juan Antonio Fafián-Labora	2020	43%
Study summary Full title Small Extracellular Vesicles Have GST Activity and Ameliorate Senescence-Related Tissue Damage All authors Juan Antonio Fafián-Labora, Jose Antonio Rodríguez-Navarro, Ana O'Loghlen Journal Cell metab Abstract Aging is a process of cellular and tissue dysfunction characterized by different hallmarks, includin (show more...)Aging is a process of cellular and tissue dysfunction characterized by different hallmarks, including cellular senescence. However, there is proof that certain features of aging and senescence can be ameliorated. Here, we provide evidence that small extracellular vesicles (sEVs) isolated from primary fibroblasts of young human donors ameliorate certain biomarkers of senescence in cells derived from old and Hutchinson-Gilford progeria syndrome donors. Importantly, sEVs from young cells ameliorate senescence in a variety of tissues in old mice. Mechanistically, we identified sEVs to have intrinsic glutathione-S-transferase activity partially due to the high levels of expression of the glutathione-related protein (GSTM2). Transfection of recombinant GSTM2 into sEVs derived from old fibroblasts restores their antioxidant capacity. sEVs increase the levels of reduced glutathione and decrease oxidative stress and lipid peroxidation both in vivo and in vitro. Altogether, our data provide an indication of the potential of sEVs as regenerative therapy in aging. (hide) EV-METRIC 43% (81st 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 Commercial method Protein markers EV: None non-EV: None Proteomics no EV density (g/ml) 1.074-1.106 Show all info Study aim Function Sample Species Homo sapiens Sample Type Cell culture supernatant EV-producing cells primary human foreskin fibroblasts EV-harvesting Medium EV-depleted medium Preparation of EDS overnight (16h) at >= 100,000g 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) 80 Pelleting: rotor type T-865 Pelleting: speed (g) 100000 Wash: volume per pellet (ml) 15 Wash: time (min) 80 Wash: Rotor Type T-865 Wash: speed (g) 100000 Density gradient Type Discontinuous Number of initial discontinuous layers 3 Lowest density fraction 10% Highest density fraction 60% Total gradient volume, incl. sample (mL) 5.5 Sample volume (mL) 1.5 Orientation Bottom-up Rotor type T-865 Speed (g) 100000 Duration (min) 720 Fraction volume (mL) 0.7 Fraction processing Centrifugation Pelleting: volume per fraction 15 Pelleting: duration (min) 80 Pelleting: rotor type T-865 Pelleting: speed (g) 100000 Pelleting-wash: volume per pellet (mL) 15 Pelleting-wash: duration (min) 80 Pelleting-wash: speed (g) T-865 Commercial kit qEV EV-subtype Distinction between multiple subtypes Size Used subtypes >200 nm Characterization: Protein analysis None Protein Concentration Method Not determined Characterization: Lipid analysis No Characterization: Particle analysis NTA Report type Mean Reported size (nm) >200 EV concentration Yes Particle yield Number of particles of starting sample E08-E09

	EV200036	10/16	Homo sapiens	primary human foreskin fibroblasts	DG (d)(U)C qEV	Juan Antonio Fafián-Labora	2020	43%
Study summary Full title Small Extracellular Vesicles Have GST Activity and Ameliorate Senescence-Related Tissue Damage All authors Juan Antonio Fafián-Labora, Jose Antonio Rodríguez-Navarro, Ana O'Loghlen Journal Cell metab Abstract Aging is a process of cellular and tissue dysfunction characterized by different hallmarks, includin (show more...)Aging is a process of cellular and tissue dysfunction characterized by different hallmarks, including cellular senescence. However, there is proof that certain features of aging and senescence can be ameliorated. Here, we provide evidence that small extracellular vesicles (sEVs) isolated from primary fibroblasts of young human donors ameliorate certain biomarkers of senescence in cells derived from old and Hutchinson-Gilford progeria syndrome donors. Importantly, sEVs from young cells ameliorate senescence in a variety of tissues in old mice. Mechanistically, we identified sEVs to have intrinsic glutathione-S-transferase activity partially due to the high levels of expression of the glutathione-related protein (GSTM2). Transfection of recombinant GSTM2 into sEVs derived from old fibroblasts restores their antioxidant capacity. sEVs increase the levels of reduced glutathione and decrease oxidative stress and lipid peroxidation both in vivo and in vitro. Altogether, our data provide an indication of the potential of sEVs as regenerative therapy in aging. (hide) EV-METRIC 43% (81st 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 iRas 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 Commercial method Protein markers EV: None non-EV: None Proteomics no EV density (g/ml) 1.074-1.106 Show all info Study aim Function Sample Species Homo sapiens Sample Type Cell culture supernatant EV-producing cells primary human foreskin fibroblasts EV-harvesting Medium EV-depleted medium Preparation of EDS overnight (16h) at >= 100,000g 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) 80 Pelleting: rotor type T-865 Pelleting: speed (g) 100000 Wash: volume per pellet (ml) 15 Wash: time (min) 80 Wash: Rotor Type T-865 Wash: speed (g) 100000 Density gradient Type Discontinuous Number of initial discontinuous layers 3 Lowest density fraction 10% Highest density fraction 60% Total gradient volume, incl. sample (mL) 5.5 Sample volume (mL) 1.5 Orientation Bottom-up Rotor type T-865 Speed (g) 100000 Duration (min) 720 Fraction volume (mL) 0.7 Fraction processing Centrifugation Pelleting: volume per fraction 15 Pelleting: duration (min) 80 Pelleting: rotor type T-865 Pelleting: speed (g) 100000 Pelleting-wash: volume per pellet (mL) 15 Pelleting-wash: duration (min) 80 Pelleting-wash: speed (g) T-865 Commercial kit qEV EV-subtype Distinction between multiple subtypes Size Used subtypes >200 nm Characterization: Protein analysis None Protein Concentration Method Not determined Characterization: Lipid analysis No Characterization: Particle analysis NTA Report type Mean Reported size (nm) >200 EV concentration Yes Particle yield Number of particles of starting sample E08-E09

	EV200036	12/16	Homo sapiens	primary human foreskin fibroblasts	DG (d)(U)C qEV	Juan Antonio Fafián-Labora	2020	43%
Study summary Full title Small Extracellular Vesicles Have GST Activity and Ameliorate Senescence-Related Tissue Damage All authors Juan Antonio Fafián-Labora, Jose Antonio Rodríguez-Navarro, Ana O'Loghlen Journal Cell metab Abstract Aging is a process of cellular and tissue dysfunction characterized by different hallmarks, includin (show more...)Aging is a process of cellular and tissue dysfunction characterized by different hallmarks, including cellular senescence. However, there is proof that certain features of aging and senescence can be ameliorated. Here, we provide evidence that small extracellular vesicles (sEVs) isolated from primary fibroblasts of young human donors ameliorate certain biomarkers of senescence in cells derived from old and Hutchinson-Gilford progeria syndrome donors. Importantly, sEVs from young cells ameliorate senescence in a variety of tissues in old mice. Mechanistically, we identified sEVs to have intrinsic glutathione-S-transferase activity partially due to the high levels of expression of the glutathione-related protein (GSTM2). Transfection of recombinant GSTM2 into sEVs derived from old fibroblasts restores their antioxidant capacity. sEVs increase the levels of reduced glutathione and decrease oxidative stress and lipid peroxidation both in vivo and in vitro. Altogether, our data provide an indication of the potential of sEVs as regenerative therapy in aging. (hide) EV-METRIC 43% (81st 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 iRas+GSTM2 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 Commercial method Protein markers EV: None non-EV: None Proteomics no EV density (g/ml) 1.074-1.106 Show all info Study aim Function Sample Species Homo sapiens Sample Type Cell culture supernatant EV-producing cells primary human foreskin fibroblasts EV-harvesting Medium EV-depleted medium Preparation of EDS overnight (16h) at >= 100,000g 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) 80 Pelleting: rotor type T-865 Pelleting: speed (g) 100000 Wash: volume per pellet (ml) 15 Wash: time (min) 80 Wash: Rotor Type T-865 Wash: speed (g) 100000 Density gradient Type Discontinuous Number of initial discontinuous layers 3 Lowest density fraction 10% Highest density fraction 60% Total gradient volume, incl. sample (mL) 5.5 Sample volume (mL) 1.5 Orientation Bottom-up Rotor type T-865 Speed (g) 100000 Duration (min) 720 Fraction volume (mL) 0.7 Fraction processing Centrifugation Pelleting: volume per fraction 15 Pelleting: duration (min) 80 Pelleting: rotor type T-865 Pelleting: speed (g) 100000 Pelleting-wash: volume per pellet (mL) 15 Pelleting-wash: duration (min) 80 Pelleting-wash: speed (g) T-865 Commercial kit qEV EV-subtype Distinction between multiple subtypes Size Used subtypes >200 nm Characterization: Protein analysis None Protein Concentration Method Not determined Characterization: Lipid analysis No Characterization: Particle analysis NTA Report type Mean Reported size (nm) >200 EV concentration Yes Particle yield Number of particles of starting sample E08-E09

	EV200036	14/16	Homo sapiens	primary human foreskin fibroblasts	DG (d)(U)C qEV	Juan Antonio Fafián-Labora	2020	43%
Study summary Full title Small Extracellular Vesicles Have GST Activity and Ameliorate Senescence-Related Tissue Damage All authors Juan Antonio Fafián-Labora, Jose Antonio Rodríguez-Navarro, Ana O'Loghlen Journal Cell metab Abstract Aging is a process of cellular and tissue dysfunction characterized by different hallmarks, includin (show more...)Aging is a process of cellular and tissue dysfunction characterized by different hallmarks, including cellular senescence. However, there is proof that certain features of aging and senescence can be ameliorated. Here, we provide evidence that small extracellular vesicles (sEVs) isolated from primary fibroblasts of young human donors ameliorate certain biomarkers of senescence in cells derived from old and Hutchinson-Gilford progeria syndrome donors. Importantly, sEVs from young cells ameliorate senescence in a variety of tissues in old mice. Mechanistically, we identified sEVs to have intrinsic glutathione-S-transferase activity partially due to the high levels of expression of the glutathione-related protein (GSTM2). Transfection of recombinant GSTM2 into sEVs derived from old fibroblasts restores their antioxidant capacity. sEVs increase the levels of reduced glutathione and decrease oxidative stress and lipid peroxidation both in vivo and in vitro. Altogether, our data provide an indication of the potential of sEVs as regenerative therapy in aging. (hide) EV-METRIC 43% (81st 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 iC+GSTM2 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 Commercial method Protein markers EV: None non-EV: None Proteomics no EV density (g/ml) 1.074-1.106 Show all info Study aim Function Sample Species Homo sapiens Sample Type Cell culture supernatant EV-producing cells primary human foreskin fibroblasts EV-harvesting Medium EV-depleted medium Preparation of EDS overnight (16h) at >= 100,000g 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) 80 Pelleting: rotor type T-865 Pelleting: speed (g) 100000 Wash: volume per pellet (ml) 15 Wash: time (min) 80 Wash: Rotor Type T-865 Wash: speed (g) 100000 Density gradient Type Discontinuous Number of initial discontinuous layers 3 Lowest density fraction 10% Highest density fraction 60% Total gradient volume, incl. sample (mL) 5.5 Sample volume (mL) 1.5 Orientation Bottom-up Rotor type T-865 Speed (g) 100000 Duration (min) 720 Fraction volume (mL) 0.7 Fraction processing Centrifugation Pelleting: volume per fraction 15 Pelleting: duration (min) 80 Pelleting: rotor type T-865 Pelleting: speed (g) 100000 Pelleting-wash: volume per pellet (mL) 15 Pelleting-wash: duration (min) 80 Pelleting-wash: speed (g) T-865 Commercial kit qEV EV-subtype Distinction between multiple subtypes Size Used subtypes >200 nm Characterization: Protein analysis None Protein Concentration Method Not determined Characterization: Lipid analysis No Characterization: Particle analysis NTA Report type Mean Reported size (nm) >200 EV concentration Yes Particle yield Number of particles of starting sample E08-E09

	EV200036	16/16	Homo sapiens	primary human foreskin fibroblasts	DG (d)(U)C qEV	Juan Antonio Fafián-Labora	2020	43%
Study summary Full title Small Extracellular Vesicles Have GST Activity and Ameliorate Senescence-Related Tissue Damage All authors Juan Antonio Fafián-Labora, Jose Antonio Rodríguez-Navarro, Ana O'Loghlen Journal Cell metab Abstract Aging is a process of cellular and tissue dysfunction characterized by different hallmarks, includin (show more...)Aging is a process of cellular and tissue dysfunction characterized by different hallmarks, including cellular senescence. However, there is proof that certain features of aging and senescence can be ameliorated. Here, we provide evidence that small extracellular vesicles (sEVs) isolated from primary fibroblasts of young human donors ameliorate certain biomarkers of senescence in cells derived from old and Hutchinson-Gilford progeria syndrome donors. Importantly, sEVs from young cells ameliorate senescence in a variety of tissues in old mice. Mechanistically, we identified sEVs to have intrinsic glutathione-S-transferase activity partially due to the high levels of expression of the glutathione-related protein (GSTM2). Transfection of recombinant GSTM2 into sEVs derived from old fibroblasts restores their antioxidant capacity. sEVs increase the levels of reduced glutathione and decrease oxidative stress and lipid peroxidation both in vivo and in vitro. Altogether, our data provide an indication of the potential of sEVs as regenerative therapy in aging. (hide) EV-METRIC 43% (81st 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 mCherry-CD63 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 Commercial method Protein markers EV: None non-EV: None Proteomics no EV density (g/ml) 1.074-1.106 Show all info Study aim Function Sample Species Homo sapiens Sample Type Cell culture supernatant EV-producing cells primary human foreskin fibroblasts EV-harvesting Medium EV-depleted medium Preparation of EDS overnight (16h) at >= 100,000g 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) 80 Pelleting: rotor type T-865 Pelleting: speed (g) 100000 Wash: volume per pellet (ml) 15 Wash: time (min) 80 Wash: Rotor Type T-865 Wash: speed (g) 100000 Density gradient Type Discontinuous Number of initial discontinuous layers 3 Lowest density fraction 10% Highest density fraction 60% Total gradient volume, incl. sample (mL) 5.5 Sample volume (mL) 1.5 Orientation Bottom-up Rotor type T-865 Speed (g) 100000 Duration (min) 720 Fraction volume (mL) 0.7 Fraction processing Centrifugation Pelleting: volume per fraction 15 Pelleting: duration (min) 80 Pelleting: rotor type T-865 Pelleting: speed (g) 100000 Pelleting-wash: volume per pellet (mL) 15 Pelleting-wash: duration (min) 80 Pelleting-wash: speed (g) T-865 Commercial kit qEV EV-subtype Distinction between multiple subtypes Size Used subtypes >200 nm Characterization: Protein analysis None Protein Concentration Method Not determined Characterization: Lipid analysis No Characterization: Particle analysis NTA Report type Mean Reported size (nm) >200 EV concentration Yes Particle yield Number of particles of starting sample E08-E09

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EV-TRACK ID	EV200036
species	Homo sapiens
sample type	Cell culture
cell type	human skin primary fibroblasts	human skin primary fibroblasts	human skin primary fibroblasts	primary human foreskin fibroblasts	primary human foreskin fibroblasts	primary human foreskin fibroblasts	primary human foreskin fibroblasts	primary human foreskin fibroblasts	human skin primary fibroblasts	human skin primary fibroblasts	human skin primary fibroblasts	primary human foreskin fibroblasts	primary human foreskin fibroblasts	primary human foreskin fibroblasts	primary human foreskin fibroblasts	primary human foreskin fibroblasts
condition	Young donors	Old donors	Progeria patients	Control condition	iRas	iRas GSTM2	iC GSTM2	mCherry-CD63	Young donors	Old donors	Progeria patients	Control condition	iRas	iRas GSTM2	iC GSTM2	mCherry-CD63
separation protocol	DG (d)(U)C qEV	DG (d)(U)C qEV	DG (d)(U)C qEV	DG (d)(U)C qEV	DG (d)(U)C qEV	DG (d)(U)C qEV	DG (d)(U)C qEV	DG (d)(U)C qEV	DG (d)(U)C qEV	DG (d)(U)C qEV	DG (d)(U)C qEV	DG (d)(U)C qEV	DG (d)(U)C qEV	DG (d)(U)C qEV	DG (d)(U)C qEV	DG (d)(U)C qEV
EV subtype	<200 nm	<200 nm	<200 nm	<200 nm	<200 nm	<200 nm	<200 nm	<200 nm	>200 nm	>200 nm	>200 nm	>200 nm	>200 nm	>200 nm	>200 nm	>200 nm
Exp. nr.	1	3	5	7	9	11	13	15	2	4	6	8	10	12	14	16
EV-METRIC %	78	78	56	56	56	56	56	56	43	43	43	43	43	43	43	43

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