Evaluation of Oxidative Stress and Mitophagy during Adipogenic Differentiation of Adipose-Derived Stem Cells Isolated from Equine Metabolic Syndrome (EMS) Horses

Figure 7

Mitochondrial dynamics and clearance in ASC_CTRL and ASC_EMS cultured under adipogenic conditions. Using fluorescence staining, cells’ cytoskeleton (f-actin) and mitochondrial net were visualized (a). Moreover, a TEM microscope allowed for a deeper assessment of mitochondrial morphology (b). Mitochondria from ASC_CTRL presented typical, elongated morphology, while ASC_EMS cells were characterized by mitochondrial aberrations including membrane and cristae raptures. Moreover, mitochondrial dynamics was assessed by RT-PCR as the expression of Mnf (c) and Fis (d) genes was investigated. However, no differences were observed between groups. Similarly, no differences in PINK expression were noted (e). To evaluate the amount of Parkin protein, immunofluorescence staining and RT-PCR were performed. Representative photographs obtained from a confocal microscope indicated increased Parkin accumulation in ASC_EMS (f). Those data were also confirmed by the analysis of Parkin mRNA levels, as it was significantly upregulated in ASC_EMS (g). Results are expressed as mean ± SD. Scale bar 250 μm.

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