Journal: Journal of Extracellular Vesicles
Article Title: Role of Collection Media on the Biological Activity of Extracellular Vesicles From hTERT‐Immortalised Mesenchymal Stromal Cells
doi: 10.1002/jev2.70298
Figure Lengend Snippet: Evaluation of the biological potential of BM‐MSC/TERT292‐derived EVs enriched from different collection media. (A) Evaluation of the dose‐dependency of the anti‐inflammatory activity of BM‐MSC/TERT292 EVs enriched from different collection media on NO secretion by LPS‐stimulated BV‐2 cells. LPS‐stimulated BV‐2 cells were treated with increasing EV concentrations ranging from 1E+08 to 2E+09 p/mL. 2.5 µM Dexamethasone (Dexa, green dotted line) was included as positive control. Data are represented as mean ± SD and data were normalised on the LPS‐stimulated BV‐2 cells (set at 100%, red dotted line). (B) Assessment of the anti‐fibrosis effect of BM‐MSC/TERT292 EVs enriched from different collection media on α‐SMA induction by TGF‐β1‐stimulated fHDF/TERT166 cells. 20 mM HEPES was included as negative control and 2 µM PP2 was included as positive control. TGF‐β1‐stimulated fHDF/TERT166 cells were treated with a fixed EV concentration of 1E+09 p/mL. (C) Quantification of the wound closure of HUVEC/TERT2 cells upon BM‐MSC/TERT292‐derived EVs treatment. Wound closure was measured at 0, 16 and 24 h after the removal of culture insert and treatment. 20 mM HEPES was included as negative control and complete expansion medium (Medium + FBS) was included as positive control. HUVEC/TERT2 cells were treated with a fixed EV concentration of 1E+09 p/mL. Data are represented as mean ± SD. One‐way Anova was used for the anti‐inflammatory and anti‐fibrosis assay; data were normalised on the LPS‐stimulated BV‐2 cells and TGF‐β1‐stimulated fHDF/TERT166 cells respectively (set at 100%). Two‐way Anova was used for the Wound Healing assay. Significance is shown as * p < 0.05, ** p < 0.01, *** p < 0.001, **** p < 0.0001 ( n = 4 technical replicates for the anti‐fibrosis and anti‐inflammatory assays, n = 3 biological replicates for the wound‐healing assay). (D) Representative microscope images of the HUVEC/TERT2 cells' wound closure 24 h following culture insert removal and treatment (photos were taken at 100× lens magnification).
Article Snippet: Human foreskin fibroblasts fHDF/TERT166 (Evercyte GmbH) were cultured in DMEM/F12 (PAN Biotech), supplemented with 10% FBS (PAN Biotech) and 100 μg/mL G418 (InvivoGen).
Techniques: Derivative Assay, Activity Assay, Positive Control, Negative Control, Concentration Assay, Wound Healing Assay, Microscopy