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Related Experiment Video

Updated: Jun 6, 2026

An In Vitro 3D Model and Computational Pipeline to Quantify the Vasculogenic Potential of iPSC-Derived Endothelial Progenitors
06:36

An In Vitro 3D Model and Computational Pipeline to Quantify the Vasculogenic Potential of iPSC-Derived Endothelial Progenitors

Published on: May 13, 2019

Quantification of circulating endothelial progenitor cells using the modified ISHAGE protocol.

Caroline Schmidt-Lucke1, Stephan Fichtlscherer, Alexandra Aicher

  • 1Department of Molecular Cardiology, Internal Medicine III, J.W. Goethe University, Frankfurt, Germany. caroline.schmidt-lucke@charite.de

Plos One
|November 13, 2010
PubMed
Summary

A new protocol accurately quantifies endothelial progenitor cells (EPCs), revealing they are lower in cardiovascular disease patients. This method identifies the CD45(dim) fraction as crucial for EPC enumeration and monitoring treatment efficacy.

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Area of Science:

  • Cardiovascular Research
  • Immunology
  • Cell Biology

Background:

  • Endothelial progenitor cells (EPCs) are vital for vascular health, playing roles in regeneration and neovascularization.
  • Current methods for EPC characterization, such as flow cytometry using markers like CD34+KDR+ or CD133+KDR+, suffer from inter-laboratory variability due to a lack of standardized protocols.
  • This variability hinders accurate assessment of EPCs' prognostic value in cardiovascular diseases.

Purpose of the Study:

  • To develop and validate a novel, standardized protocol for accurate enumeration of circulating endothelial progenitor cells (EPCs).
  • To adapt the established ISHAGE protocol for hematopoietic stem cell enumeration to improve EPC quantification.
  • To assess the clinical utility of the new protocol in distinguishing EPC levels between healthy controls and patients with coronary artery disease (CAD).

Main Methods:

  • Adapted the standardized ISHAGE protocol for enumerating hematopoietic stem cells to quantify EPCs in whole blood.
  • Incubated blood samples with CD45, KDR, and CD34 antibodies, employing a sequential gating strategy.
  • Quantified CD45(dim)CD34(+)KDR(+) cells, comparing results with CD45(+)CD34(+)KDR(+) and CD45(-)CD34(+)KDR(+) populations in healthy controls and CAD patients.

Main Results:

  • The novel protocol identified significantly higher CD45(dim)CD34(+)KDR(+) EPCs in healthy controls compared to patients with CAD or acute coronary syndrome (ACS).
  • A significant inverse correlation was observed between CD45(dim)CD34(+)KDR(+) EPC levels and cardiovascular disease activity, including the number of diseased coronary arteries.
  • Atorvastatin treatment in stable CAD patients led to a significant increase specifically in the CD45(dim)CD34(+)KDR(+) EPC population.

Conclusions:

  • The newly established protocol, adapted from the ISHAGE standard, provides higher accuracy in EPC enumeration.
  • The CD45(dim) fraction is identified as the key compartment harboring EPCs.
  • The protocol confirms the inverse correlation of EPCs with disease activity and their increase during statin therapy, highlighting its clinical relevance.