Figure 1.
Gating strategy to characterize circulating endothelial cells (CECs) by surface markers.
(A) Human CEC Panel: (1) whole fresh blood; (2) select Hoechst33342+ cells; (3) CD45−/CD31+ cells; (4) CD31+/CD146+ co-expression; and (5) select CD133−/Hoechst 33342+ cells (CECs). (B) Mouse CEC Panel: (1) whole fresh blood; (2) select Hoechst 33342+ cells; (3) select CD45−/CD31+ cells; (4) CD31+/KDR+ co expression; and (5) select CD117−/Hoechst 33342+ cells (CECs).
Figure 2.
Intra-assay variability analysis of CECs in murine and human peripheral blood.
CEC counts are expressed as a percentage of peripheral blood mononuclear cells. Intra-assay variability is presented as the CV value of 8 replicates, calculated as (standard deviation/mean)×100%. (A) In 5 VEGF-treated mice (M1–5), the CV values were 9.3% (M1), 5.7% (M2), 7.0% (M3), 7.6% (M4), and 6.5% (M5). (B) In 5 AMI patients (H1–5), the CV values were 10.3% (H1), 9.6% (H2), 9.4% (H3), 8.5% (H4), and 6.9% (H5).
Figure 3.
CECs in mouse peripheral blood detected by flow cytometry and real-time PCR.
(A) CEC percentage detected by flow cytometry. (B) CEC concentration detected by real-time PCR. KDR is an endothelial cell marker. (n = 5/group, **p<0.01).
Figure 4.
Daily variability in CEC concentration.
(A) Peripheral blood samples of VEGF-treated mice were collected at the same time and the CEC concentration was determined at different time points (0, 24, 48, and 72 h). (B) Peripheral blood samples of AMI patients were collected at different time points (0, 24, 48, and 72 h) and the CEC concentrations were determined. The error bar represents the standard deviation of 4 replicates. (*p<0.05, **p<0.01).
Figure 5.
CEC concentrations in AMI patients and healthy controls.
CEC counts are expressed as the percentage of peripheral blood mononuclear cells. **p<0.01.
Table 1.
Characteristics of patients.
Table 2.
Biochemical markers and cardiac enzymes.
Figure 6.
Changes in CEC concentrations post-treatment.
CEC concentrations are expressed as the percentage of peripheral blood mononuclear cells. *p<0.05.