Enhancement of Mitochondrial Transfer by Antioxidants in Human Mesenchymal Stem Cells

Figure 1

MCA improved mitochondrial biogenesis and bioenergetics in hMSCs. Mitochondrial dehydrogenase activity (a) and mitochondrial mass (b) of hMSCs treated with or without MCA for 24 h was assessed by CCK-8 assay and flow cytometric analysis of MitoTracker Red-labeled mitochondria, respectively. (c) The cell density of hMSCs with or without MCA treatment for 24 h was assessed by counting the harvested cells and normalized to the area of the culture flask. (d) The mitochondria of the hMSCs with or without MCA treatment were labeled with MitoTracker Red and subjected to confocal microscopy. The mitochondria were classified using MicroP software according to their morphology: small globules, linear tubules, loops, branched tubules, and twisted tubules. Three major types of mitochondria were quantified: globular, tubular, and branched ((e), upper panel), as well as their total length ((e), lower panel). (f) Mitochondrial bioenergetics of the indicated cells was determined by oxygen consumption rate (OCR) using Oroboros Oxygraph-2k analyzer. The MCA consisted of 2 mM NAC and 0.2 mM AAP. The concentrations of NAC and AAP were 2 mM and 0.2 mM, respectively, in monotreatments. OCR measured at basal respiration (upper left), maximal respiration (upper right), ATP-coupled respiration (lower left), and spare capacity (lower right) were shown. , , , as compared to the control (Ctrl); n.s., not significant.

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