Figure 1.
Flow cytometry strategy to detect CD109+CECs and CD146+CECs.
Fluorescence Minus One for each fluorescence is reported.
Figure 2.
Real-time quantitative PCR analysis comparing changes in gene expression between CD109+ cells and CD146+ cells.
Fold changes of down- or up-regulated genes are expressed comparing cDNA from CD109+ to CD146+ cells as reference cells. To ensure biological reproducibility, blood for cDNA collection were obtained from 8 different donors.
Figure 3.
Flow cytometry strategy to detect CD109+CEC and CD146+CEC phenotype.
After exclusion of debris (A) and selection of CD45− (B), nucleated (Syto16+) and CD31+ cells (C), CEC were identified as positive for CD109 or CD146 (E). (D): negative control. CD109+ CECs and CD146+CECs were evaluated by flow cytometry for the expression of CD34, CD117, CD90 and CD13.
Figure 4.
Expression of Ulex Europeaus Lectin.on CD109+CECs and CD146+CECs+ (4A–B1).
Epcam staining (4C–C1) confirmed that Epithelial Cells even if present in the DNA+CD45− cell compartment, are negative for the expression of CD31 present only on endothelial cells.
Figure 5.
Flow cytometry strategy to detect Ac-LDL uptake.
After dublets (A) and debris (B) exclusion, Ac-LDL+ cells were detected (C). Among these cells, CD45+CD31+ and CD45−CD31+ cells were detected (D). CD45+CD31+ were also positive for CD14 (monocytes,E) and CD45−CD31+ were also positive for CD109+ (endothelial cells, F). Ac-LDL+ cells were nucleated (Syto16+, F). C1, D1, E1 and F1 are the negative controls.
Table 1.
Patients characteristics.
Table 2.
Baseline values of CD109+ CECs and CD146+ CECs.
Table 3.
Viable CD109+ CECs and CD146+CECs before and after treatment.