Journal: Journal of Extracellular Biology
Article Title: iMSC‐Derived Extracellular Vesicles Improve Atopic Dermatitis by Augmenting Skin Barrier Integrity and Inhibiting Inflammation, Pruritus and Th2 Immune Responses
doi: 10.1002/jex2.70067
Figure Lengend Snippet: Characterization of IFN‐γ‐iMSCs and IFN‐γ‐iMSC‐EVs. (a) Flow cytometry analysis of IFN‐γ‐iMSCs. The reactivities of IFN‐γ‐iMSCs against positive (CD105, CD73 and CD90) or negative (CD45, CD31 and CD34) markers of MSCs were evaluated. (b) Representative image of IDO1 protein expression in iMSCs and IFN‐γ‐iMSCs by immunocytochemistry analysis. Green and blue denote cells stained with anti‐IDO1 antibody and DAPI, respectively. (c) Immunoblot analysis of IDO1 protein levels in iMSCs and IFN‐γ‐iMSCs. n = 3. Data are presented as mean ± SE. ** p < 0.01 versus Control. (d) Scheme of the IFN‐γ‐iMSC‐EVs manufacturing. IFN‐γ‐iMSC‐EVs were isolated from the IFN‐γ‐iMSCs culture medium using ultracentrifugation. (e) Morphology of IFN‐γ‐iMSC‐EVs under cryo‐TEM. Scale bar = 200 nm. (f) Size distribution of IFN‐γ‐iMSC‐EVs shown by NTA. The average size is 134.6 nm. (g) Western blot analyses for markers of extracellular vesicles (CD9 and CD63) or cellular organelles (Cytochrome C and Histone H3) in IFN‐γ‐iMSCs and IFN‐γ‐iMSC‐EVs. (h) Expression analysis of EV markers (CD63 and CD81) in IFN‐γ‐iMSC‐EVs by flow cytometry.
Article Snippet: To verify the expression of typical MSC surface markers on IFN‐γ‐primed iMSCs (IFN‐γ‐iMSCs), they were stained with antibodies against CD73 APC, CD105 PE, CD45 FITC and CD34 APC (eBioscience, Waltham, MA, USA), as well as CD90 APC‐Cy7 (BioLegend, San Diego, CA, USA).
Techniques: Flow Cytometry, Expressing, Immunocytochemistry, Staining, Western Blot, Control, Isolation