Extracellular Vesicles are Associated with Human Donor Kidney Characteristics 5 129 6.39E5 ± 4.67E5 objects/mL. For CD81 and CD31 double-positive events, we observed >99% reduction of fluorescent events after detergent treatment, ~950 / 1130 / 1100 – fold difference in objects/mL at each time point of sample drawing compared to T0 (2.2E4 ± 1.9E4 objects/mL), and an isotype background of 3.28E4 ± 1.53E4 objects/ mL (Figure 3D). We then analyzed the relative abundance of both double-positive EV populations with respect to the total CD81+ EVs detected. We observed that 6.8% of CD81+ EVs expressed the endothelial cell marker CD31, and only 1.7% of CD81+ EVs was found to express the common leukocyte antigen CD45 (Figure 3E). The far majority of CD81+ EVs detected (91.5%) were found to not express either of the measured markers. In summary, these data show that leukocyte and endothelial-derived EVs are released during NMP. Surprisingly, the majority of CD81+ EVs did not bear either of the studied markers.
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