Figure 5

Flow cytometry analysis of circulating EC after transplantation. Flow Cytometry Analysis of Circulating EC after Transplantation. Example of a flow cytometric analysis of circulating EC in peripheral blood. The depicted data show a blood sample from an EPC-injected lung transplant recipient 12 h post reperfusion. Whole blood was stained with CD42d-FITC to exclude platelet aggregates and with CD45-FITC to exclude leukocytes, as well as CD146-PE (P1H12) as second fluorescence. Circulating EC are defined as CD146 positive and negative for CD45 and CD42d. The localization of that cell group in forward and side scatter is shown in Panel A as the green cell group. Panel A and B contain furthermore as a red group a standard dose of control beads in order to quantify cells. Panel A shows the forward and side scatter characteristics of the blood sample. In panel B the latex beads which are used as internal quantification standard (gate Beads) and the CD45 and CD42a negative population (gate R2) are gated. In panel C the CD45 and CD42a negative population of R2 is analyzed for their expression of the endothelial cell marker P1H12. Cells were counted as circulating endothelial cells when they were included in the depicted gate CEC.