MSCs were immunophenotypically characterised by flow cytometry on a BD FACS Canto II (BD Biosciences). As the negative control we used 106 unstained MSCs. Test samples contained 106 MSCs with PE Mouse Anti-Human CD73, APC Mouse Anti-Human CD105, PerCP-Cy5.5 Mouse Anti-Human CD90, FITC Mouse Anti-Human CD31 and PerCP-Cy7 Mouse Anti-Human CD45 (all BD Biosciences). In addition, FACS analysis was used to determine the effect of NMP on the survival of MSCs during perfusion without a kidney with pre-labelled MSCs. For this purpose, hourly perfusate samples were taken. The MSCs could be identified by their fluorescence emission at two different wavelengths as a result of the PKH26 and Q-tracker 655 pre-labelling.
Flow cytometry was also performed on perfusate samples and enzymatically disrupted kidney biopsies from the experiments with pre-labelled MSCs with a kidney in the perfusion circuit. Of each sample, 1 million counts were obtained and analysed through flow cytometry.
Flow cytometry was also performed on perfusate samples and enzymatically disrupted kidney biopsies from the experiments with pre-labelled MSCs with a kidney in the perfusion circuit. Of each sample, 1 million counts were obtained and analysed through flow cytometry.