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(a–c) Characterization and quantification of human-bone marrow mesenchymal stem cell (hBM-MSC)-derived extracellular vesicles (EVs). (a) Nanosight Tracking analyses (NTA) showing the size distribution pattern of EVs. (b) Western blots of hBM-MSCs marker (GM130) and EVs markers (ALIX, CD9, CD81). “Wash” lane represents the negative control. Each image comes from different blots. (c) Representative image of EVs using transmission electron microscopy (TEM). Scale bar 100 nm. (d–f) In vivo tracking of ExoGlow-labeled EVs demonstrated that EVs reach the brain and accumulate near the stroke injury 6 hours after administration at 2 dps. (d) Timeline of intranasal single dose treatment of ExoGlow-labeled EVs (~2.4 x 10 9 EVs in 200 μl), starting 2 dps and collecting the tissue 6 hours after. (e) Representative coronal section after a single intranasal administration of ExoGlow-labeled EVs (~2.4 x 10 9 EVs in 200 μl) at 2 dps. Hoechst indicates cell nuclei. Scale bars 4 mm and 100 μm. (f) Quantification of the average size of ExoGlow+ particles in the ipsilateral and contralateral hemispheres. Data was analyzed using a paired t-test (*p<0.05). Bars show mean +/− SD. Each symbol represents the average size of ExoGlow+ particles from all ROIs in each hemisphere (5 ROIs/hemisphere in each rat) of a single rat ( n = 4 ).

Journal: PLOS ONE

Article Title: Recovery after human bone marrow mesenchymal stem cells (hBM-MSCs)-derived extracellular vesicles (EVs) treatment in post-MCAO rats requires repeated handling

doi: 10.1371/journal.pone.0312298

Figure Lengend Snippet: (a–c) Characterization and quantification of human-bone marrow mesenchymal stem cell (hBM-MSC)-derived extracellular vesicles (EVs). (a) Nanosight Tracking analyses (NTA) showing the size distribution pattern of EVs. (b) Western blots of hBM-MSCs marker (GM130) and EVs markers (ALIX, CD9, CD81). “Wash” lane represents the negative control. Each image comes from different blots. (c) Representative image of EVs using transmission electron microscopy (TEM). Scale bar 100 nm. (d–f) In vivo tracking of ExoGlow-labeled EVs demonstrated that EVs reach the brain and accumulate near the stroke injury 6 hours after administration at 2 dps. (d) Timeline of intranasal single dose treatment of ExoGlow-labeled EVs (~2.4 x 10 9 EVs in 200 μl), starting 2 dps and collecting the tissue 6 hours after. (e) Representative coronal section after a single intranasal administration of ExoGlow-labeled EVs (~2.4 x 10 9 EVs in 200 μl) at 2 dps. Hoechst indicates cell nuclei. Scale bars 4 mm and 100 μm. (f) Quantification of the average size of ExoGlow+ particles in the ipsilateral and contralateral hemispheres. Data was analyzed using a paired t-test (*p<0.05). Bars show mean +/− SD. Each symbol represents the average size of ExoGlow+ particles from all ROIs in each hemisphere (5 ROIs/hemisphere in each rat) of a single rat ( n = 4 ).

Article Snippet: Human bone marrow mesenchymal stem cells (hBM-MSCs) were purchased from Celprogren (frozen vial with ~1.2 x 10 6 cells; #36094–22) and plated on human mesenchymal bone marrow stem cell culture extracellular expansion matrix pre-coated T75 Flasks (Celprogen, #E36094-21-T75) following Celprogren recommendations.

Techniques: Derivative Assay, Western Blot, Marker, Negative Control, Transmission Assay, Electron Microscopy, In Vivo, Labeling