Unit

UNIT 9.33 Identification of Endothelial Cells and Progenitor Cell Subsets in Human Peripheral Blood

  1. Myka L. Estes1,2,
  2. Julie A. Mund1,2,
  3. David A. Ingram1,2,3,
  4. Jamie Case1,2

Published Online: 1 APR 2010

DOI: 10.1002/0471142956.cy0933s52

Current Protocols in Cytometry

Current Protocols in Cytometry

How to Cite

Estes, M. L., Mund, J. A., Ingram, D. A. and Case, J. 2010. Identification of Endothelial Cells and Progenitor Cell Subsets in Human Peripheral Blood. Current Protocols in Cytometry. 52:9.33:9.33.1–9.33.11.

Author Information

  1. 1

    Department of Pediatrics, Indiana University School of Medicine, Indianapolis, Indiana

  2. 2

    Herman B. Wells Center for Pediatric Research, Indianapolis, Indiana

  3. 3

    Department of Biochemistry and Molecular Biology, Indiana University School of Medicine, Indianapolis, Indiana

Publication History

  1. Published Online: 1 APR 2010
  2. Published Print: APR 2010

Abstract

An assay for circulating cell subsets in human peripheral blood by flow cytometry is used as a biomarker to determine cardiovascular disease risk and tumor responsiveness to chemotherapy since endothelial progenitor cells (EPCs) function in vasculogenesis and angiogenesis. Despite analytical advances in polychromatic flow cytometry (PFC), conventional approaches are routinely utilized to enumerate and isolate EPCs, which has led to varied results in clinical studies, potential cellular misidentification, and thus a lack of a plausible biological explanation for how purported EPCs function. Herein, a reproducible PFC protocol is provided to identify a rare circulating endothelial colony-forming cell (ECFC) with proliferative potential, along with a population of circulating progenitor cells (CPCs) in which the ratio analysis distinguishes between healthy and disease populations. In sum, a reliable PFC protocol, which can be used to investigate the roles of human hematopoietic and endothelial elements in the growth and maintenance of the vasculature, is described. Curr. Protoc. Cytom. 52:9.33.1-9.33.11. © 2010 by John Wiley & Sons, Inc.

Keywords:

  • endothelial cells;
  • circulating endothelial cells;
  • endothelial progenitor cells;
  • circulating endothelial progenitors;
  • human peripheral blood