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. Author manuscript; available in PMC: 2024 Mar 1.
Published in final edited form as: Adv Ther (Weinh). 2022 Dec 9;6(3):2200197. doi: 10.1002/adtp.202200197

Figure 1. Vasculogenic EVs fabrication, isolation, and characterization.

Figure 1.

A) Schematic describing how engineered vasculogenic EVs are derived from primary human dermal fibroblasts after electroporation with plasmids encoding for the vasculogenic transcription factors genes ETV2, FLI1, and FOXC2 (EFF). Nanoparticle tracking analysis showing B) particle concentration and C) particle size distribution for EFF and control EVs isolated at 12, 24, and 48 hours after electroporation of the donor cells. D) CryoEM images showing single EFF and control EVs isolated 48 hours post-electroporation of the donor cells, showing their intact morphology with a defined lipid bilayer. E) Western blot characterization of EFF and control EVs showing positive expression of the EV marker, CD63 for both EV preparations. F) ETV2, FLI1, and FOXC2 (EFF) upregulation in donor cells at 12, 24, and 48 hours after electroporation compared to sham electroporated cells. G) Dynamics of transcript packing in EFF and control EVs isolated at 12, 24, and 48 hours after electroporation of donor cells. H) Transcript levels of additional vasculogenic factors (VEGF-A, VEGF-KDR, and FGF-2) packed in EFF and control EVs isolated 48 hours after electroporation of donor cells. Data represented as standard deviation Mean+/−SD. # Indicates significant difference with respect to control EVs. * # P < 0.05 (n = 3 – 6, One way ANOVA with Fisher LSD multiple comparison test).