In vitro characterization of circulating endothelial progenitor cells isolated from patients with acute coronary
Diana Campioni1, Giorgio Zauli, Stefania Gambetti
1Department of Medical Sciences, Section of Hematology, Azienda Ospedaliero-Universitaria, Arcispedale Sant'Anna, University of Ferrara, Ferrara, Italy.
Insights
This study investigated circulating endothelial progenitor cells (EPC) in acute coronary syndrome (ACS) patients, finding that while hematopoietic cells were common, true EPC/ECFC were rare. Careful characterization of EPC is crucial for future clinical applications in ACS.
Area of Science:
- Cardiovascular Research
- Cell Biology
- Hematology
Background:
- Understanding circulating endothelial progenitor cells (EPC) is limited, particularly in cardiovascular disease patients.
- This study focuses on the in vitro clonogenic capacity of EPC, also known as endothelial colony-forming cells (ECFC).
- Comparison is made between ECFC and colony-forming unit-endothelial-like cells (CFU-EC) of hematopoietic origin in acute coronary syndrome (ACS) patients.
Purpose of the Study:
- To analyze the in vitro clonogenic capacity of circulating EPC/ECFC in ACS patients.
- To compare the presence and characteristics of EPC/ECFC with CFU-EC in ACS patients.
- To lay the groundwork for future clinical studies involving autologous EPC in ACS.
Main Methods:
- Peripheral blood mononuclear cells (PBMC) from 70 ACS patients were cultured.
- Specific endothelial culture media were used for EPC/ECFC expansion.
- Phenotypic and functional characterization included FISH analysis, immunophenotyping (CD markers), and single-cell replanting assays.
Main Results:
- CFU-EC were frequently isolated (77% of patients), while EPC/ECFC were rare (13%).
- EPC/ECFC generation correlated with higher in vitro PDGF-AA release.
- Expanded EPC/ECFC showed normal diploid chromosomes, specific immunophenotypes (CD105+, CD31+, CD146+, Factor VIII+, variable CD34, CD106, CD184, negative for CD45, CD90, CD117, CD133), and clonogenic expansion capacity.
Conclusions:
- True EPC require careful characterization in the context of ACS.
- Findings highlight the rarity of functional EPC/ECFC in ACS patients.
- This research is essential for designing future clinical trials using autologous EPC in ACS.
Background:
The current understanding of the functional characteristics of circulating endothelial progenitor cells (EPC) is limited, especially in patients affected by cardiovascular diseases. In this study, we have analyzed the in vitro clonogenic capacity of circulating EPC, also known as endothelial colony-forming cells (ECFC), in patients with acute coronary syndrome (ACS), in comparison to the colony forming unit-endothelial-like cells (CFU-EC) of hematopoietic/monocytic origin.
Methodology/Principal Findings:
By culturing peripheral blood mononuclear cells (PBMC) of patients with ACS (n = 70), CFU-EC were frequently isolated (from 77% of ACS patients), while EPC/ECFC were obtained only in a small subset (13%) of PBMC samples, all harvested between 7-14 days after the acute cardiovascular event. Notably, ex-vivo generation of EPC/ECFC was correlated to a higher in vitro release of PDGF-AA by the corresponding ACS patient PBMC. By using specific endothelial culture media, EPC/ECFC displayed in vitro expansion capacity, allowing the phenotypic and functional characterization of the cells. Indeed, after expansion, EPC/ECFC exhibited a normal diploid chromosomal setting by FISH analysis and an immunophenotype characterized by: i) uniform positivity for the expression of CD105, CD31, CD146 and Factor VIII, i) variable expression of the CD34, CD106 and CD184 markers, and iii) negativity for CD45, CD90, CD117 and CD133. Of interest, in single-cell replanting assays EPC/ECFC exhibited clonogenic expansion capacity, forming secondary colonies characterized by variable proliferation capacities.
Conclusion/Significance:
Our data indicate that a careful characterization of true EPC is needed in order to design future studies in the clinical autologous setting of patients with ACS.
