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Updated: Jul 15, 2026
Phenotypic and Functional Characterization of Endothelial Colony Forming Cells Derived from Human Umbilical Cord Blood
Published on: April 13, 2012
A protocol for phenotypic detection and enumeration of circulating endothelial cells and circulating progenitor cells
Dan G Duda1, Kenneth S Cohen, David T Scadden
1Steele Laboratory for Tumor Biology, Department of Radiation Oncology, Massachusetts General Hospital, Boston, Massachusetts 02114, USA.
Insights
This study introduces a simple, cost-effective cytometry protocol to identify and count circulating endothelial cells (CECs) and circulating hematopoietic progenitor cells (CPCs) in human blood for disease research.
Area of Science:
- Biomedical Engineering
- Cell Biology
- Hematology
Background:
- Circulating endothelial cells (CECs) and circulating hematopoietic progenitor cells (CPCs) are crucial for tissue vascularization and serve as biomarkers for various diseases.
- Current research is hindered by inconsistent methodologies for CEC and CPC evaluation, leading to data interpretation challenges.
Purpose of the Study:
- To establish a standardized, reliable cytometry protocol for the phenotypic identification and enumeration of CECs and CPCs in human blood.
- To provide a platform for consistent longitudinal studies in patients with diverse pathologies.
Main Methods:
- A flow cytometry protocol utilizing four surface markers: CD31, CD34, CD133, and CD45.
- The protocol is designed for human blood, focusing on phenotypic analysis and cell enumeration within the mononuclear cell population.
Main Results:
- The proposed method enables phenotypic analysis and enumeration of CECs and CPCs.
- The protocol is efficient, taking 2-2.5 hours, and adaptable to various flow cytometry platforms.
- Expected detection rates are 0.1-6.0% for viable CECs and 0.01-0.20% for CPCs within the mononuclear cell population.
Conclusions:
- This cytometry protocol offers a standardized, accessible, and cost-effective approach for CEC and CPC analysis.
- It facilitates further biological exploration and consistent patient monitoring across different pathologies.
Abstract:
Blood circulating endothelial cells (CECs) and circulating hematopoietic progenitor cells (CPCs) represent two cell populations that are thought to play important roles in tissue vascularization. CECs and CPCs are currently studied as surrogate markers in patients for more than a dozen pathologies, including heart disease and cancer. However, data interpretation has often been difficult because of multiple definitions, methods and protocols used to evaluate and count these cells by different laboratories. Here, we propose a cytometry protocol for phenotypic identification and enumeration of CECs and CPCs in human blood using four surface markers: CD31, CD34, CD133 and CD45. This method allows further phenotypic analyses to explore the biology of these cells. In addition, it offers a platform for longitudinal studies of these cells in patients with different pathologies. The protocol is relatively simple, inexpensive and can be adapted for multiple flow cytometer types or software. The procedure should take 2-2.5 h, and is expected to detect 0.1-6.0% viable CECs and 0.01-0.20% CPCs within blood mononuclear cell population.
