Journal: Journal of Extracellular Vesicles

Article Title: Evaluation of unmodified human cell‐derived extracellular vesicle mitochondrial deoxyribonucleic acid‐based biodistribution in rodents

doi: 10.1002/jev2.12489

Figure Lengend Snippet: Quantitative range assessment of five EVs obtained from three commercial sources and one self‐isolated EVM.

Article Snippet: The quantification potentials of Expi293F EVM and HEK293 large EV from HansaBioMed, HEK293 EV‐GFPs from Sigma‐Aldrich, and BM‐MSC EV and AD‐MSC EV from STEMON were evaluated, and their quantification ranges were compared utilizing RNR1 analysis to confirm the applicability of mtDNA‐qPCR for analysing other human‐derived EVs.

Techniques:

Journal: Journal of Extracellular Vesicles

Article Title: Evaluation of unmodified human cell‐derived extracellular vesicle mitochondrial deoxyribonucleic acid‐based biodistribution in rodents

doi: 10.1002/jev2.12489

Figure Lengend Snippet: EVMs isolated from Expi293F and primer and probe performance check. (a) Nanoparticle tracking analysis was employed to unveil the size distribution and concentration of Expe293F‐EVs particles. (b) Expe293F‐EVs were visualized using transmission electron microscopy. Standard curve for detecting Expi293F‐EVMs DNA using primer and probe sets for RNR1 (c,) (d), and (e) and ND6 (f) and (g). Quantitative PCR was performed in triplicate in a single run, and mean Ct values were plotted. A Ct value of 40 was assumed when samples were not detected. Negative control samples contained mouse gDNA only and were used as NTC (red dot). The detection threshold was established as two cycles below the lower Ct value of NTC (indicated by red and dotted lines), and the corresponding number of cycles is highlighted in red. The linear approximation equations (represented by black lines) and their corresponding R 2 values are displayed. Although undetected, a dot was placed at the 40‐cycle mark for visualization. EV, extracellular vesicle; EVM, EV mimetic; NTC, non‐template controls; PCR, polymerase chain reaction.

Article Snippet: The quantification potentials of Expi293F EVM and HEK293 large EV from HansaBioMed, HEK293 EV‐GFPs from Sigma‐Aldrich, and BM‐MSC EV and AD‐MSC EV from STEMON were evaluated, and their quantification ranges were compared utilizing RNR1 analysis to confirm the applicability of mtDNA‐qPCR for analysing other human‐derived EVs.

Techniques: Isolation, Concentration Assay, Transmission Assay, Electron Microscopy, Real-time Polymerase Chain Reaction, Negative Control, Polymerase Chain Reaction

Journal: Journal of Extracellular Vesicles

Article Title: Evaluation of unmodified human cell‐derived extracellular vesicle mitochondrial deoxyribonucleic acid‐based biodistribution in rodents

doi: 10.1002/jev2.12489

Figure Lengend Snippet: Summary of validation for Expi293F EVM quantification.

Article Snippet: The quantification potentials of Expi293F EVM and HEK293 large EV from HansaBioMed, HEK293 EV‐GFPs from Sigma‐Aldrich, and BM‐MSC EV and AD‐MSC EV from STEMON were evaluated, and their quantification ranges were compared utilizing RNR1 analysis to confirm the applicability of mtDNA‐qPCR for analysing other human‐derived EVs.

Techniques: Biomarker Discovery

Journal: Journal of Extracellular Vesicles

Article Title: Evaluation of unmodified human cell‐derived extracellular vesicle mitochondrial deoxyribonucleic acid‐based biodistribution in rodents

doi: 10.1002/jev2.12489

Figure Lengend Snippet: In vivo imaging. Representative tissue whole body images from mice infused with IRDye 800CW‐labeled Expi293F EVMs. Representative images of the whole body from mice administered EVMs. Fluorescence signal analysis of vehicle control ( n = 1), EVMs‐IRDye 800CW‐labeled group ( n = 5), and free IRDye 800CW dye group ( n = 2) is shown. The fluorescence intensity reached its highest level 30 min after EV administration. The images were relatively adjusted based on the signal, and all measurements were performed using the same setting value. EVM, extracellular vesicle mimetic.

Article Snippet: The quantification potentials of Expi293F EVM and HEK293 large EV from HansaBioMed, HEK293 EV‐GFPs from Sigma‐Aldrich, and BM‐MSC EV and AD‐MSC EV from STEMON were evaluated, and their quantification ranges were compared utilizing RNR1 analysis to confirm the applicability of mtDNA‐qPCR for analysing other human‐derived EVs.

Techniques: In Vivo Imaging, Labeling, Fluorescence, Control

Journal: Journal of Extracellular Vesicles

Article Title: Evaluation of unmodified human cell‐derived extracellular vesicle mitochondrial deoxyribonucleic acid‐based biodistribution in rodents

doi: 10.1002/jev2.12489

Figure Lengend Snippet: Ex vivo imaging. Representative tissue organ images from mice infused with IRDye 800CW‐labeled Expi293F‐EVMs and comparison of SNR by organ. Representative images of dissected organs from mice administered EVMs. Fluorescence signal analysis of vehicle control ( n = 1), EVMs‐IRDye 800CW‐labelled group ( n = 5), and free IRDye 800CW dye group ( n = 2) is shown. The fluorescence intensity reached its highest level 30 min after EV administration. The images were relatively adjusted based on the signal, and all measurements were performed using the same setting value. The SNR analysis was performed for each organ, including the liver, lungs, kidneys, spleen, digestive tract, and heart. The SNR values were calculated based on the signal intensity and background noise levels observed in the images. Sequential IRDye 800CW signal in the organs from free dye only (b) or Expi293F EVMs‐administrated mice (c). Mean ± standard deviation of SNR obtained from two (free dye only) or five (Expi293F EVMs) are plotted against days after inoculation. EVM, extracellular vesicle mimetic; SNR, signal‐to‐noise ratio.

Article Snippet: The quantification potentials of Expi293F EVM and HEK293 large EV from HansaBioMed, HEK293 EV‐GFPs from Sigma‐Aldrich, and BM‐MSC EV and AD‐MSC EV from STEMON were evaluated, and their quantification ranges were compared utilizing RNR1 analysis to confirm the applicability of mtDNA‐qPCR for analysing other human‐derived EVs.

Techniques: Ex Vivo, Imaging, Labeling, Comparison, Fluorescence, Control, Standard Deviation

Journal: Journal of Extracellular Vesicles

Article Title: Evaluation of unmodified human cell‐derived extracellular vesicle mitochondrial deoxyribonucleic acid‐based biodistribution in rodents

doi: 10.1002/jev2.12489

Figure Lengend Snippet: Biodistribution of Expi293F‐EVMs in nude BALB/c mice after tail vein injection. The biodistribution analysis of Expi293F‐EVMs in nude BALB/c mice following intravenous administration. Nude BALB/c mice were injected via the tail vein with 1 × 10 11 Expi293F EVMs, vehicle control (PBS), or free dye control. Tissue samples, including the heart (a), lungs (b), liver (c), kidneys (d), spleen (e), and gastrointestinal tract, were collected at time points 0.5, 3, and 24 h post‐administration of Expi293F EVs. Biodistribution data in each tissue are presented as EVs/µg mouse DNA. All data is presented as mean ± standard deviation ( n = 5 animals per group). In all the graphs, the red line indicates the lower limit of the quantification threshold (1 × 10 2 EVs/µg mouse DNA). DNA, deoxyribonucleic acid; EVM, extracellular vesicle mimetics; PBS, phosphate‐buffered saline.

Article Snippet: The quantification potentials of Expi293F EVM and HEK293 large EV from HansaBioMed, HEK293 EV‐GFPs from Sigma‐Aldrich, and BM‐MSC EV and AD‐MSC EV from STEMON were evaluated, and their quantification ranges were compared utilizing RNR1 analysis to confirm the applicability of mtDNA‐qPCR for analysing other human‐derived EVs.

Techniques: Injection, Control, Standard Deviation, Saline

Journal: Journal of Extracellular Vesicles

Article Title: Evaluation of unmodified human cell‐derived extracellular vesicle mitochondrial deoxyribonucleic acid‐based biodistribution in rodents

doi: 10.1002/jev2.12489

Figure Lengend Snippet: Comparison of Expi293F‐EVMs biodistribution in different organs at various time points. The figure presents a comparison of EV biodistribution in different organs at three time points: 0.5, 3, and 24 h. The biodistribution in each organ is represented by the (a), (b), and (c), expressed as EVs per µg of mouse DNA. Additionally, the concentrations in organs (d), (e), and (f) were normalized using organ weight to account for differences in concentration owing to varying organ sizes. All data is presented as mean ± standard deviation ( n = 5 animals per group). DNA, deoxyribonucleic acid; EVM, extracellular vesicle mimetics.

Article Snippet: The quantification potentials of Expi293F EVM and HEK293 large EV from HansaBioMed, HEK293 EV‐GFPs from Sigma‐Aldrich, and BM‐MSC EV and AD‐MSC EV from STEMON were evaluated, and their quantification ranges were compared utilizing RNR1 analysis to confirm the applicability of mtDNA‐qPCR for analysing other human‐derived EVs.

Techniques: Comparison, Concentration Assay, Standard Deviation

Journal: Cell

Article Title: Asymmetric and non-stoichiometric glycoprotein recognition by two distinct antibodies results in broad protection against ebolaviruses

doi: 10.1016/j.cell.2022.02.023

Figure Lengend Snippet: Key resources table

Article Snippet: Drosophila S2, ExpiCHO-S and Expi293F cell lines were obtained from ThermoFisher Scientific.

Techniques: Labeling, Virus, Luciferase, Recombinant, Blocking Assay, Electron Microscopy, Enzyme-linked Immunosorbent Assay, Expressing, Transfection, Modification, Saline, TaqMan Assay, Plasmid Preparation, Software, High Content Screening, Microscopy, Real-time Polymerase Chain Reaction