Journal: International Journal of Molecular Sciences
Article Title: Mitochondrial Metabolism and EV Cargo of Endothelial Cells Is Affected in Presence of EVs Derived from MSCs on Which HIF Is Activated
doi: 10.3390/ijms24066002
Figure Lengend Snippet: DFO-sEV production and HUVEC treatment. ( a ) Representative image of DFO-sEVs at TEM. sEVs appear with the typical bilayer cup-shaped membrane structure. ( b ) Particle size distribution and concentration of DFO-sEVs analyzed by tunable resistive pulse sensing: mean diameter of 90 ± 30.9 nm, mode of 73 nm, and particle size distribution of D10 67, D50 82, and D90 121. The average concentration was 1.33 × 10 9 particles/mL. ( c ) Flow cytometry of DFO-sEVs showing positivity to surface markers: CD81 and CD63 (DFO-sEVs in red, vehicle in gray). ( d ) Representative image of the uptake of PKH67-labeled green fluorescent DFO-sEVs (left) and of negative control, i.e., PKH67-labeled PBS, (right) after 24 h of incubation. Nuclei stained with Hoechst 33342 (blue). In red area, 2× magnification of adjacent green area.
Article Snippet: Bead-bound sEVs were washed twice with an Assay Buffer (0.1% BSA in PBS), and then labeled with 20 μL of mouse anti-human CD81-PE monoclonal antibody (BD Pharmingen™, BD Biosciences, San Jose, CA, USA) or 5 μL of mouse anti-human CD63-PE monoclonal antibody (eBioscience, San Diego, CA, USA).
Techniques: Concentration Assay, Tunable Resistive Pulse Sensing, Flow Cytometry, Labeling, Negative Control, Incubation, Staining