Journal: Journal of Extracellular Vesicles
Article Title: Functional diversity of apoptotic vesicle subpopulations from bone marrow mesenchymal stem cells in tissue regeneration
doi: 10.1002/jev2.12434
Figure Lengend Snippet: Induction of apoptosis in rat MSCs and characterization of apoptotic vesicle subtypes. (a) Schematic representation of the differential centrifugation method used to isolate apoptotic vesicle subtypes, apoBD and apoSEV, from apoptotic MSCs. (b) TEM images capturing the vesicle morphology of apoBD and apoSEV subtypes. Scale bar, 500 nm (c) Size distribution of apoptotic vesicles assessed through DLS, demonstrating the distinct size ranges of apoBD and apoSEV, and their average particle size was counted. (d) Exposure of Annexin V on the surface of isolated apoptotic vesicles, indicating their apoptotic nature. Scale bar, 5 μm (e) apoSEV and apoBD were co‐stained with Annexin V and PKH26, with both markers expressed on the vesicle membrane surfaces. Scale bar, 5 μm (f) Positive expression of apoptotic vesicles observed via nano‐flow cytometry. (g) Western blot analysis demonstrated the presence of Caspase‐3/Cleaved Caspase‐3 in MSC, apoSEV and apoBD. (h) PKH26‐labeled apoSEV and apoBD were both efficiently engulfed by MSCs. DLS, dynamic light scattering; MSCs, mesenchymal stem cells; TEM, transmission electron microscopy.
Article Snippet: Human bone marrow mesenchymal stem cells (h‐BMSCs) were purchased from cyagen Company in China and cultured in complete medium with the same ratios as the r‐BMSCs described above under the same conditions, and cells with less than five generations of passages were used for the subsequent studies.
Techniques: Centrifugation, Isolation, Staining, Membrane, Expressing, Flow Cytometry, Western Blot, Labeling, Transmission Assay, Electron Microscopy
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