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Details	EV-TRACK ID	Experiment nr.	Species	Sample type	Separation protocol	First author	Year	EV-METRIC
	EV200030	1/2	Homo sapiens	oral mucosa lamina propria progenitor cells	(d)(U)C Other/ ExoSpin UF Filtration DG	Knight R	2022	100%
Study summary Full title Oral Progenitor Cell Line-Derived Small Extracellular Vesicles as a Treatment for Preferential Wound Healing Outcome. All authors Knight R, Board-Davies E, Brown H, Clayton A, Davis T, Karatas B, Burston J, Tabi Z, Falcon-Perez JM, Paisey S, Stephens P Journal Stem Cells Transl Med Abstract Scar formation during wound repair can be devastating for affected individuals. Our group previously (show more...)Scar formation during wound repair can be devastating for affected individuals. Our group previously documented the therapeutic potential of novel progenitor cell populations from the non-scarring buccal mucosa. These Oral Mucosa Lamina Propria-Progenitor Cells (OMLP-PCs) are multipotent, immunosuppressive, and antibacterial. Small extracellular vesicles (sEVs) may play important roles in stem cell-mediated repair in varied settings/ hence, we investigated sEVs from this source for wound repair. We created an hTERT immortalized OMLP-PC line (OMLP-PCL) and confirmed retention of morphology, lineage plasticity, surface markers, and functional properties. sEVs isolated from OMLP-PCL were analyzed by nanoparticle tracking analysis, Cryo-EM and flow cytometry. Compared to bone marrow-derived mesenchymal stromal cells (BM-MSC) sEVs, OMLP-PCL sEVs were more potent at driving wound healing functions, including cell proliferation and wound repopulation and downregulated myofibroblast formation. A reduced scarring potential was further demonstrated in a preclinical in vivo model. Manipulation of OMLP-PCL sEVs may provide novel options for non-scarring wound healing in clinical settings. (hide) EV-METRIC 100% (99th percentile of all experiments on the same sample type) Reported Not reported Not applicable EV-enriched proteins Protein analysis: analysis of three or more EV-enriched proteins non EV-enriched protein Protein analysis: assessment of a non-EV-enriched protein qualitative and quantitative analysis Particle analysis: implementation of both qualitative and quantitative methods. For the quantitative method, the reporting of measured EV concentration is expected. electron microscopy images Particle analysis: inclusion of a widefield and close-up electron microscopy image density gradient Separation method: density gradient, at least as validation of results attributed to EVs EV density Separation method: reporting of obtained EV density ultracentrifugation specifics Separation method: reporting of g-forces, duration and rotor type of ultracentrifugation steps antibody specifics Protein analysis: antibody clone/reference number and dilution lysate preparation Protein analysis: lysis buffer composition Study data Sample type Cell culture supernatant Sample origin Control condition Focus vesicles Other/ small extracellular vesicles 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 Commercial method Ultrafiltration Filtration Density gradient Protein markers EV: CD81/ CD63/ CD9 non-EV: CD105/ CD90/ CD166 Proteomics no Show all info Study aim Function Sample Species Homo sapiens Sample Type Cell culture supernatant EV-producing cells oral mucosa lamina propria progenitor cells EV-harvesting Medium EV-depleted medium Preparation of EDS >=18h at >= 100,000g Separation Method (Differential) (ultra)centrifugation dUC: centrifugation steps Below or equal to 800 g Pelleting performed No Density gradient Only used for validation of main results Yes Type Continuous Lowest density fraction 0.2M Highest density fraction 2.5M Total gradient volume, incl. sample (mL) 5 Sample volume (mL) 0.2 Orientation Bottom-up Rotor type MLS-50 Speed (g) 200000 Duration (min) 16 Fraction volume (mL) 0.3 Fraction processing None Filtration steps 0.22µm or 0.2µm Ultra filtration Cut-off size (kDa) 100 Membrane type Polyethersulfone (PES) Commercial kit Other/ ExoSpin Characterization: Protein analysis Protein Concentration Method microBCA Flow cytometry aspecific beads Antibody details provided? No Detected EV-associated proteins CD9/ CD63/ CD81 Not detected contaminants CD90/ CD105/ CD166 Flow cytometry specific beads Antibody details provided? Yes Antibody dilution provided? No Characterization: Lipid analysis No Characterization: Particle analysis NTA Report type Modus Reported size (nm) 94 EV concentration Yes Particle yield as number of particles per milliliter of starting sample: 7.7E+12 EM EM-type Cryo-EM Image type Close-up, Wide-field

	EV200030	2/2	Homo sapiens	bone marrow derived mesenchymal stromal cells	(d)(U)C Other/ ExoSpin UF Filtration DG	Knight R	2022	100%
Study summary Full title Oral Progenitor Cell Line-Derived Small Extracellular Vesicles as a Treatment for Preferential Wound Healing Outcome. All authors Knight R, Board-Davies E, Brown H, Clayton A, Davis T, Karatas B, Burston J, Tabi Z, Falcon-Perez JM, Paisey S, Stephens P Journal Stem Cells Transl Med Abstract Scar formation during wound repair can be devastating for affected individuals. Our group previously (show more...)Scar formation during wound repair can be devastating for affected individuals. Our group previously documented the therapeutic potential of novel progenitor cell populations from the non-scarring buccal mucosa. These Oral Mucosa Lamina Propria-Progenitor Cells (OMLP-PCs) are multipotent, immunosuppressive, and antibacterial. Small extracellular vesicles (sEVs) may play important roles in stem cell-mediated repair in varied settings/ hence, we investigated sEVs from this source for wound repair. We created an hTERT immortalized OMLP-PC line (OMLP-PCL) and confirmed retention of morphology, lineage plasticity, surface markers, and functional properties. sEVs isolated from OMLP-PCL were analyzed by nanoparticle tracking analysis, Cryo-EM and flow cytometry. Compared to bone marrow-derived mesenchymal stromal cells (BM-MSC) sEVs, OMLP-PCL sEVs were more potent at driving wound healing functions, including cell proliferation and wound repopulation and downregulated myofibroblast formation. A reduced scarring potential was further demonstrated in a preclinical in vivo model. Manipulation of OMLP-PCL sEVs may provide novel options for non-scarring wound healing in clinical settings. (hide) EV-METRIC 100% (99th percentile of all experiments on the same sample type) Reported Not reported Not applicable EV-enriched proteins Protein analysis: analysis of three or more EV-enriched proteins non EV-enriched protein Protein analysis: assessment of a non-EV-enriched protein qualitative and quantitative analysis Particle analysis: implementation of both qualitative and quantitative methods. For the quantitative method, the reporting of measured EV concentration is expected. electron microscopy images Particle analysis: inclusion of a widefield and close-up electron microscopy image density gradient Separation method: density gradient, at least as validation of results attributed to EVs EV density Separation method: reporting of obtained EV density ultracentrifugation specifics Separation method: reporting of g-forces, duration and rotor type of ultracentrifugation steps antibody specifics Protein analysis: antibody clone/reference number and dilution lysate preparation Protein analysis: lysis buffer composition Study data Sample type Cell culture supernatant Sample origin Control condition Focus vesicles Other/ small extracellular vesicles 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 Commercial method Ultrafiltration Filtration Density gradient Protein markers EV: CD81/ CD63/ CD9 non-EV: None Proteomics no Show all info Study aim Function Sample Species Homo sapiens Sample Type Cell culture supernatant EV-producing cells bone marrow derived mesenchymal stromal cells EV-harvesting Medium EV-depleted medium Preparation of EDS >=18h at >= 100,000g Separation Method (Differential) (ultra)centrifugation dUC: centrifugation steps Below or equal to 800 g Pelleting performed No Density gradient Only used for validation of main results Yes Type Continuous Lowest density fraction 0.2M Highest density fraction 2.5M Total gradient volume, incl. sample (mL) 5 Sample volume (mL) 0.2 Orientation Bottom-up Rotor type MLS-50 Speed (g) 200000 Duration (min) 16 Fraction volume (mL) 0.3 Fraction processing None Filtration steps 0.22µm or 0.2µm Ultra filtration Cut-off size (kDa) 100 Membrane type Polyethersulfone (PES) Commercial kit Other/ ExoSpin Characterization: Protein analysis Protein Concentration Method microBCA Flow cytometry aspecific beads Antibody details provided? No Detected EV-associated proteins CD9/ CD63/ CD81 Not detected contaminants CD90/ CD105/ CD166 Flow cytometry specific beads Antibody details provided? Yes Antibody dilution provided? No Characterization: Lipid analysis No Characterization: Particle analysis NTA Report type Modus Reported size (nm) 99.5 EV concentration Yes Particle yield as number of particles per milliliter of starting sample: 4.27E+12 EM EM-type Cryo-EM Image type Close-up, Wide-field

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EV-TRACK ID	EV200030
species	Homo sapiens
sample type	Cell culture
cell type	oral mucosa lamina propria progenitor cells	bone marrow derived mesenchymal stromal cells
condition	Control condition	Control condition
separation protocol	dUC/ Other/ ExoSpin/ Ultrafiltration/ Filtration/ Density gradient	dUC/ Other/ ExoSpin/ Ultrafiltration/ Filtration/ Density gradient
Exp. nr.	1	2
EV-METRIC %	100	100