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Details	EV-TRACK ID	Experiment nr.	Species	Sample type	Separation protocol	First author	Year	EV-METRIC
	EV200089	1/1	Homo sapiens	red blood cells	(d)(U)C DC Filtration	Zhang, Gensheng; Huang, Xiaofa	2020	67%
Study summary Full title Extracellular vesicles: Natural liver‐accumulating drug delivery vehicles for the treatment of liver diseases All authors Gensheng Zhang, Xiaofang Huang, Huiqing Xiu, Yan Sun, Jiming Chen, Guoping Cheng, Zhengbo Song, Yanmei Peng, Yingying Shen, Jianli Wang, Zhijian Cai Journal J Extracell Vesicles Abstract Extracellular vesicles (EVs) are excellent potential vectors for the delivery of therapeutic drugs. (show more...)Extracellular vesicles (EVs) are excellent potential vectors for the delivery of therapeutic drugs. However, issues with biological safety and disease targeting substantially limit their clinical application. EVs from red blood cells (RBC‐EVs) are potential drug delivery vehicles because of their unique biological safety. Here, we demonstrated that EVs, including RBC‐EVs, show natural liver accumulation. Mechanistically, the liver environment induces macrophages to phagocytize RBC‐EVs in a C1q‐dependent manner. RBC‐EVs loaded with antisense oligonucleotides of microRNA‐155 showed macrophage‐dependent protective effects against acute liver failure (ALF) in a mouse model. These RBC‐EVs were also effective in treatment of ALF. Furthermore, compared to routine doses of doxorubicin and sorafenib (SRF), RBC‐EVs loaded with doxorubicin or SRF showed enhanced therapeutic effects on a murine model of orthotopic liver cancer through a mechanism dependent on macrophages. Importantly, drug‐loaded RBC‐EVs showed no systemic toxicity at therapeutically effective doses, whereas routine doses of doxorubicin and SRF showed obvious toxicity. Thus, drug‐loaded RBC‐EVs hold high potential for clinical applications in the treatment of liver disease therapy. (hide) EV-METRIC 67% (50th 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 red blood cells 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 (d)(U)C DC Filtration Protein markers EV: TSG101/ HSP70/ Alix non-EV: ARF6 Proteomics no Show all info Study aim Function/Biomarker/Mechanism of uptake/transfer Sample Species Homo sapiens Sample Type red blood cells Separation Method (Differential) (ultra)centrifugation dUC: centrifugation steps Below or equal to 800 g Between 800 g and 10,000 g Between 100,000 g and 150,000 g Pelleting performed Yes Pelleting: time(min) 70 Pelleting: rotor type SW 32 Ti Pelleting: speed (g) 100000 Wash: volume per pellet (ml) 35 Wash: time (min) 70 Wash: Rotor Type SW 32 Ti Wash: speed (g) 100000 Filtration steps 0.45µm > x > 0.22µm, 0.22µm or 0.2µm Density cushion Density medium Sucrose Characterization: Protein analysis Protein Concentration Method BCA Western Blot Antibody details provided? Yes Antibody dilution provided? Yes Lysis buffer provided? Yes Detected EV-associated proteins TSG101/ HSP70/ Alix Not detected contaminants ARF6 Characterization: Lipid analysis No Characterization: Particle analysis NTA Report type Other Reported size (nm) 154+/-51 EV concentration Yes Particle yield Yes, as number of particles per milliliter of starting sample 1.84E+12 EM EM-type Transmission-EM Image type Close-up Report size (nm) 150-200

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EV-TRACK ID	EV200089
species	Homo sapiens
sample type	red blood cells
condition	Control condition
separation protocol	(d)(U)C DC Filtration
Exp. nr.	1
EV-METRIC %	67