Fig. 3
Estimation of the rescue of mitochondrial mass, metabolic activity, and mortality of damaged PBMCs using PAMM MitoCeption. PBMCs were isolated, exposed to UVR (3 min = 72 mJ/cm2), and treated with 3 allogeneic mitochondria doses (1,25-2,5–5,0 μg of mitochondria). a, b Mitochondria mass (n = 7, 7 PBMC donors exposed to UVR and PAMM of 3 donors). MFI of PBMCs labelled with MitoTracker Green® was analyzed by flow cytometry. a 1 h after exposure to UVR, 1 h after MitoCeption, wash, and 30` incubation with MitoTracker® Green. b After 1 h exposure to UVR, 1 h MitoCeption, wash and 18 h in culture. Analysis, the sample/control-average transformation was used, Anderson-Darling normality test was applied. Un-paired, ANOVA test and Student’s t-test are shown in the figure with an alpha-value of 0.05 (*p < 0.05, **p < 0.01, ***p < 0.001, ****p < 0.0001) to observe statistically significant differences. c, d Metabolic activity measured by MTT (n = 5, 5 PBMC donors exposed to UVR and PAMM of 3 donors). MAV were measured by spectrophotometry of PBMCs treated with MTT after UVR exposure and MitoCeption at the early and late time points. c 1 h after exposure to UVR, 1 h of MitoCeption, wash and 2 h of incubation with MTT. d 1 h after exposure to UVR, 1 h of MitoCeption, wash, 18 h in culture and 2 h of incubation with MTT. Analysis: the sample/control-average transformation was used, Anderson-Darling normality test was applied. Un-paired, ANOVA test and Student’s t-test are shown in the figure with an alpha-value of 0.05 (*p < 0.05, **p < 0.01, ***p < 0.001, ****p < 0.0001) to observe statistically significant differences. e, f Mortality of PBMCs caused by UVR (n = 5, 5 PBMC donors exposed to UVR and PAMM of 3 donors), the percentage was estimated by counting dead cells positive for Trypan Blue staining and dividing that number by viable cells. e Early time point measured 1 h after exposure to UVR, 1 h MitoCeption and wash f Late time point 1 h after exposure to UVR, 1 h MitoCeption, wash and 18 h in culture. Analysis: Anderson-Darling normality test was applied to the resulting data. Un-paired, ANOVA test and Student’s t-test are shown in the figure with an alpha-value of 0.05 (*p < 0.05, **p < 0.01, ***p < 0.001, ****p < 0.0001) to observe statistically significant differences