Fig. 6

Effects of mitochondria transfer on BMSC metabolism. The extracellular flux analysis of OXPHOS activity in mitochondria-recipient BMSCs versus control, including a measurements of oxygen consumption rates (OCR, pmols O2/min) throughout the entire detection process, and b–g calculations of mean OCR at different stages correlated to basal OCR (b), ATP production (c), proton leak OCR (d), maximum respiration (e), spare respiratory capacity (f), and non-mitochondrial respiration (g). Measurement of ATP production (h) by ATP luminescent detection assay. i Effects of ATPase inhibitor oligomycin on ATP production of recipient BMSCs. Representative micrographs of fluorescently labeled cells (j), quantitative results of fluorescent cell count (k), and changes in CCK8 absorbance readings (l) after treating mitochondria-recipient BMSCs with 5 or 10 μg/mL of oligomycin for 12 h. Effects of 5 or 10 μg/mL oligomycin treatment on the osteogenic differentiation potential of mitochondria-recipient BMSCs, as assessed by ALP activity (m), mRNA expression levels of Runx2 (n), and BMP2 (o) after 4 days of osteogenic induction. Effects of 5 or 10 μg/mL oligomycin treatment on the migration capacity of recipient BMSCs, as evaluated by the scratch wound healing assay (p) and the Cell-IQ single cell migration speed test (q). Results are presented as the mean ± SEM (n = 3). One-way ANOVA with Tukey’s post hoc test was used to assess statistical significance (*P < 0.05, **P < 0.01, ***P < 0.001, ****P < 0.0001)