A six-colour flow cytometric method for simultaneous detection of cell phenotype and apoptosis of circulating endothelial cells

S. Mariucci, B. Rovati, S. Chatzileontiadou, K. Bencardino, M. Manzoni, S. Delfanti, M. Danova

Research output: Contribution to journalArticlepeer-review

Abstract

Blood circulating endothelial cells (CECs), with their resting and activated subsets, (rCECs and aCECs) and circulating progenitors cells (CEPs) are two extremely rare cell populations that are important in tissue vascularization. Their number and function are modulated in diseases involving vascular injury, such as human tumours. Although a consensus on the phenotypic definition of endothelial cells, as well as on the optimal enumeration technique, is still lacking, the number of clinical studies based on assessment of these cells is rapidly expanding, as well as the analytical methods employed. The present study aimed to develop a rapid and sensitive flow cytometric method of quantifying and characterizing CECs (with both their subsets and the apoptotic fraction) and CEPs. We analysed peripheral blood samples from 21 subjects with a six-colour flow cytometric approach allowing detection of the cell phenotype of CECs and CEPs using a monoclonal antibodies panel and a dedicated gating strategy. Apoptotic CECs were detected with Annexin V and dead cells with 7-amino-actinomycin D staining. The described technique proved to be a new, reliable, tool increasing our knowledge of the biology of CECs and CEPs and can readily be applied in the study of many pathological conditions characterized by endothelial damage.

Original languageEnglish
Pages (from-to)433-438
Number of pages6
JournalScandinavian Journal of Clinical and Laboratory Investigation
Volume69
Issue number3
DOIs
Publication statusPublished - May 2009

Keywords

  • Annexin V
  • Flow cytometry
  • Immunophenotype
  • Peripheral blood
  • Rare events analysis

ASJC Scopus subject areas

  • Clinical Biochemistry

Fingerprint Dive into the research topics of 'A six-colour flow cytometric method for simultaneous detection of cell phenotype and apoptosis of circulating endothelial cells'. Together they form a unique fingerprint.

Cite this