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Details	EV-TRACK ID	Experiment nr.	Species	Sample type	Separation protocol	First author	Year	EV-METRIC
	EV230257	1/1	Lactobacillus plantarum	APsulloc 331	(d)(U)C Filtration DG	Kim H	2020	71%
Study summary Full title Comparative Lipidomic Analysis of Extracellular Vesicles Derived from APsulloc 331261 Living in Green Tea Leaves Using Liquid Chromatography-Mass Spectrometry. All authors Kim H, Kim M, Myoung K, Kim W, Ko J, Kim KP, Cho EG Journal Int J Mol Sci Abstract is a popular probiotic species due to its safe and beneficial effects on humans/ therefore, novel (show more...)is a popular probiotic species due to its safe and beneficial effects on humans/ therefore, novel strains have been isolated and identified from various dietary products. Given that bacteria-derived extracellular vesicles (EVs) have been considered as efficient carriers of bioactive materials and shown to evoke cellular responses effectively, -derived EVs are expected to efficiently elicit health benefits. Herein, we identified APsulloc 331261 living in green tea leaves and isolated EVs from the culture medium. We performed quantitative lipidomic analysis of APsulloc 331261 derived EVs (LEVs) using liquid chromatography-mass spectrometry. In comparison to APsulloc 331261, in LEVs, 67 of 320 identified lipid species were significantly increased and 19 species were decreased. In particular, lysophosphatidylserine(18:4) and phosphatidylcholine(32:2) were critically increased, showing over 21-fold enrichment in LEVs. In addition, there was a notable difference between LEVs and the parent cells in the composition of phospholipids. Our results suggest that the lipidomic profile of bacteria-derived EVs is different from that of the parent cells in phospholipid content and composition. Given that lipids are important components of EVs, quantitative and comparative analyses of EV lipids may improve our understanding of vesicle biogenesis and lipid-mediated intercellular communication within or between living organisms. (hide) EV-METRIC 71% (96th percentile of all experiments on the same sample type) Reported Not reported Not applicable EV-enriched proteins Protein analysis: analysis of three or more EV-enriched proteins non EV-enriched protein Protein analysis: assessment of a non-EV-enriched protein qualitative and quantitative analysis Particle analysis: implementation of both qualitative and quantitative methods. For the quantitative method, the reporting of measured EV concentration is expected. electron microscopy images Particle analysis: inclusion of a widefield and close-up electron microscopy image density gradient Separation method: density gradient, at least as validation of results attributed to EVs EV density Separation method: reporting of obtained EV density ultracentrifugation specifics Separation method: reporting of g-forces, duration and rotor type of ultracentrifugation steps antibody specifics Protein analysis: antibody clone/reference number and dilution lysate preparation Protein analysis: lysis buffer composition Study data Sample type Cell culture supernatant Sample origin Control condition Focus vesicles extracellular vesicle Separation protocol Separation protocol Gives a short, non-chronological overview of the different steps of the separation protocol. dUC = (Differential) (ultra)centrifugation DG = density gradient UF = ultrafiltration SEC = size-exclusion chromatography IAF = immuno-affinity capture (Differential) (ultra)centrifugation Filtration Density gradient Protein markers EV: None non-EV: None Proteomics no Show all info Study aim Identification of content (omics approaches) Sample Species Lactobacillus plantarum Sample Type Cell culture supernatant EV-producing cells APsulloc 331 EV-harvesting Medium Serum free medium Cell count 2.7E9 Separation Method (Differential) (ultra)centrifugation dUC: centrifugation steps Between 800 g and 10,000 g Between 10,000 g and 50,000 g Equal to or above 150,000 g Pelleting performed Yes Pelleting: rotor type SW 41 Ti Pelleting: speed (g) 150000 Density gradient Only used for validation of main results Yes Type Discontinuous Lowest density fraction 20% Highest density fraction 50% Orientation Bottom-up Speed (g) 200000 Duration (min) 120 Fraction processing Centrifugation Pelleting: volume per fraction 60 Pelleting: duration (min) 180 Pelleting: rotor type SW 41 Ti Pelleting: speed (g) 150000 Filtration steps 0.45µm > x > 0.22µm, Characterization: Protein analysis None Protein Concentration Method Bradford Characterization: Lipid analysis Yes Characterization: Particle analysis DLS Report type Mean Reported size (nm) 93 Used for determining EV concentration? Yes TRPS Report type Mean Reported size (nm) 93 EV concentration Yes Particle yield other:/ per mg of protein EM EM-type Cryo-EM Image type Close-up, Wide-field Report size (nm) 20-200

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EV-TRACK ID	EV230257
species	Lactobacillus plantarum
sample type	Cell culture
cell type	APsulloc 331
condition	Control condition
separation protocol	dUC/ Filtration/ Density gradient
Exp. nr.	1
EV-METRIC %	71