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Fig. 1 | Stem Cell Research & Therapy

Fig. 1

From: Mesenchymal stem cells improve redox homeostasis and mitochondrial respiration in fibroblast cell lines with pathogenic MT-ND3 and MT-ND6 variants

Fig. 1

Transfer of mitochondria and mitochondrial DNA from MSCs to fibroblasts. A Representative image of MSCs with GFP-tagged mitochondrial Cox8a and fibroblasts with BFP-tagged nuclear-located LMNB. To visualise cell structure, cells were additionally stained with Alexa Fluor 594 phalloidin (Thermo Scientific). B Quantitative flow cytometry analysis of mitochondrial transfer between fibroblasts and mesenchymal stem cells (MSCs). Mitochondrial transfer rate was measured and quantified after 72 h of co-culturing. MSCs were either co-cultured with healthy control fibroblasts (Control+) or patient fibroblasts carrying either the MT-ND6 variant (MT-ND6+), the m.10191T>C variant in the MT-ND3 gene (MT-ND3a+) or the m.10197G>A variant in the MT-ND3 gene (MT-ND3b+). The percentage (%) of fibroblasts positive for Cox8a-GFP and LMNB-RFP in relation to all LMNB-RFP positive cells are shown. Data are displayed as mean and error bars indicate standard error of the mean (SEM). *p < 0.05. C mtDNA sequencing of hypervariable regions in control and patient fibroblasts after co-culturing with MSCs. Bases at specific nucleotide positions indicated as position in the entire mitochondrial genome are shown in mesenchymal stem cells (MSC) as well as in control fibroblasts (Control) and fibroblasts carrying variants in either MT-ND6 (MT-ND6) or in MT-ND3 (m.10191T>C—MT-ND3a; m.10197G>A—MT-ND3b). Control and patient fibroblasts after 72 h hours of co-culturing with MSCs and subsequent removal of MSCs are indicated by “+ ” (e.g. “MT-ND3a+”). MSCs-treated fibroblasts mainly show changes of mtDNA in hypervariable region II (HVRII). Treated fibroblasts contain double peaks in the chromatograms (D) that vary in height. Arrows (C) indicate which peak is bigger, and further capital letters are used to indicate comparable peak heights, while small letters are used to indicate smaller peaks. Purple letters represent MSC mtDNA sequence. D Representative segments of chromatograms of sequenced regions of untreated and MSC-treated (+) control and patient fibroblasts after Sanger sequencing. Treated fibroblasts show second peaks that derive from MSC mtDNA. Nucleotide positions with two peaks are highlighted by red (position 16114 of the mtDNA) or purple box (position 16126 of the mtDNA)

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