Chapter 5 140 previously published calibration data 23. Lower and upper gating area cut-offs were defined as 677 – 112,201 ERF for BV421; 35.40 – 3776 ERF for CFSE; 206 – 14,770 ERF for PE; and 6.40 – 123 ERF for APC. Statistical Analysis Statistical analysis was performed using R version 4.0.2 and RStudio (RStudio Team (2016). RStudio: Integrated Development for R. RStudio, Inc., Boston, MA URL http://www.rstudio.com/.) version 1.1.463. Statistical significance between EV concentrations and binary data was determined through two-sided t-tests, 95% CI with unpaired data. Linear correlations with continuous variables were examined using the Pearson correlation method. R2 values ≥ 0.6 and P values < 0.05 were considered significant. ACKNOWLEDGMENTS The authors would like to thank the members of the PROPER Consortium (PROlonged ex-vivo normothermic machine PERfusion for kidney regeneration) for setting up the study, provision of the normothermic machine perfusion samples and input in research design: Prof. Dr. Ian Alwayn, Prof. Dr. Rutger Ploeg, Dr. Dorottya de Vries, Dr. Volkert Huurman, Jason Doppenberg and Asel Arykbaeva from the Leiden University Medical Center, Prof. Dr. Henri Leuvenink, Dr. Robert Pol, Dr. Cyril Moers, Tim Hamelink, Veerle Lantinga and Leonie van Leeuwen from University Medical Center Groningen and Dr. Robert Minnee from the Erasmus Medical Center, Rotterdam, the Netherlands. Additionally, the authors would like to thank Manou van Alphen for her assistance with NMP sample measurements with IFCM. AUTHOR CONTRIBUTIONS W.W.W. participated in the research design, performance of the research, data analysis, article drafting and is the corresponding author. A.A. performed the normothermic machine perfusion of kidneys and provided perfusate samples. I.P.J.A. orchestrated the prolonged normothermic machine perfusion study design. C.C.B. participated in the research design. R.C. provided clinical expertise. M.J.H., K.B. participated in the research design and writing of the manuscript. All authors reviewed the manuscript and approved its final version.
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