Journal: Cells
Article Title: Transfer of Cardiac Mitochondria Improves the Therapeutic Efficacy of Mesenchymal Stem Cells in a Preclinical Model of Ischemic Heart Disease.
doi: 10.3390/cells12040582
Figure Lengend Snippet: Figure 2. Cardiac mitochondria are internalized by MSCs through dynamin-dependent, clathrin- mediated endocytosis. (A) Representative confocal microscopy pictures of WGA-stained MSCs after 24 h of incubation with MitoTracker Green-labeled cardiac mitochondria at the Mito 3 concentration in the absence or presence of dynasore. Scale bar: 5 µm. (B) Flow cytometry quantification of Mito- Tracker Green-labeled cardiac mitochondria by MSCs following 24 h of exposure in the presence or absence of dynasore (n = 4). (C) Relative Ki67 mRNA levels in cardiac mitochondria-preconditioned MSCs in the presence or absence of dynasore in reference to their respective controls (n = 4). (D) Relative VEGF and HGF mRNA levels in cardiac mitochondria-preconditioned MSCs in the presence or absence of dynasore in reference to their respective controls (n = 4). (E) Relative mRNA levels of CXCL1, CXCL5, CXCL6, IL11, IL33 and LIF in cardiac mitochondria-preconditioned MSCs in the presence or absence of dynasore in reference to their respective controls (n = 4). (F) Relative mRNA levels of MMP1, MMP9 and MMP14 (n =4) and (G) collagenase activity (n = 10) in cardiac mitochondria-preconditioned MSCs in the presence or absence of dynasore in reference to their respective controls. One-way ANOVA with Dunn’s multiple comparisons test in (B–F). One-way ANOVA with Tukey’s multiple comparisons test in (G). * p < 0.05, ** p < 0.01, *** p < 0.001. Each dot represents an independent experiment. Bar graphs represent mean values ± SD.
Article Snippet: To characterize the endocytosis process by which MSCs internalize cardiac mitochondria, human MSCs were exposed to cardiac mitochondria previously labeled with MitoTracker Green FM (40 nM, Invitrogen, Waltham, MA, USA, Cat#M7514) in the presence of the dynamin-dependent, clathrin-mediated endocytosis inhibitor dynasore (50mM, Santa Cruz Biotechnology, Dallas, TX, USA, Cat#sc-202592).
Techniques: Confocal Microscopy, Staining, Incubation, Labeling, Concentration Assay, Flow Cytometry, Activity Assay
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