Evaluation of mobilized peripheral blood CD34+ cells from patients with severe coronary artery disease as a source of endothelial progenitor cells

Abba Zubair, Sunita Malik, Athena Paulsen, Masakazu Ishikawa, Christopher McCoy, Peter X. Adams, David Amrani, Marco Costa

Research output: Contribution to journalArticle

14 Citations (Scopus)

Abstract

Background aims. The distinction between hematopoietic stem cells (HSC) and endothelial progenitor cells (EPC) is poorly defined. Co-expression of CD34 antigen with vascular endothelial growth factor (VEGF) receptor (VEGFR2) is currently used to define EPC (1). Methods. We evaluated the phenotypic and genomic characteristics of peripheral blood-derived CD34+ cells in 22 granulocytecolony-stimulating factor (G-CSF)-mobilized patients with severe coronary artery disease and assessed the influence of cell selection and storage on CD34+ cell characteristics. Results. The median CD34+ cell contents in the products before and after enrichment with the Isolex 300i Magnetic Cell Selection System were 0.2% and 82.5%, respectively. Cell-cycle analysis showed that 80% of CD34+ cells were in G0 stage; 70% of the isolated CD34+ cells co-expressed CD133, a marker for more immature progenitors. However, less than 5% of the isolated CD34+ cells co-expressed the notch receptor Jagged-1 (CD339) and only 2% of the isolated CD34+ population were positive for VEGFR2 (CD309). Molecular assessment of the isolated CD34+ cells demonstrated extremely low expression of VEGFR2 and endothelial nitric oxide synthase (eNOS) and high expression of VEGF-A. Overnight storage at 4°C did not significantly affect CD34+ cell counts and viability. Storage in liquid nitrogen for 7 weeks did not affect the percentage of CD34+ cells but was associated with a 26% drop in cell viability. Conclusions. We have demonstrated that the majority of isolated CD34+ cells consist of immature and quiescent cells that lack prototypic markers of EPC. High VEGF-A gene expression might be one of the mechanisms for CD34+ cell-induced angiogenesis.

Original languageEnglish (US)
Pages (from-to)178-189
Number of pages12
JournalCytotherapy
Volume12
Issue number2
DOIs
StatePublished - 2010

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Coronary Artery Disease
Blood Cells
Endothelial Progenitor Cells
Vascular Endothelial Growth Factor A
Cell Survival
CD34 Antigens
Notch Receptors
Vascular Endothelial Growth Factor Receptor
Nitric Oxide Synthase Type III
Hematopoietic Stem Cells
Cell Cycle
Nitrogen
Cell Count
Gene Expression

Keywords

  • Coronary artery disease
  • Endothelial progenitor cells
  • Stem cell mobilization
  • Transplant

ASJC Scopus subject areas

  • Immunology
  • Immunology and Allergy
  • Molecular Medicine

Cite this

Evaluation of mobilized peripheral blood CD34+ cells from patients with severe coronary artery disease as a source of endothelial progenitor cells. / Zubair, Abba; Malik, Sunita; Paulsen, Athena; Ishikawa, Masakazu; McCoy, Christopher; Adams, Peter X.; Amrani, David; Costa, Marco.

In: Cytotherapy, Vol. 12, No. 2, 2010, p. 178-189.

Research output: Contribution to journalArticle

Zubair, Abba ; Malik, Sunita ; Paulsen, Athena ; Ishikawa, Masakazu ; McCoy, Christopher ; Adams, Peter X. ; Amrani, David ; Costa, Marco. / Evaluation of mobilized peripheral blood CD34+ cells from patients with severe coronary artery disease as a source of endothelial progenitor cells. In: Cytotherapy. 2010 ; Vol. 12, No. 2. pp. 178-189.
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AU - Malik, Sunita

AU - Paulsen, Athena

AU - Ishikawa, Masakazu

AU - McCoy, Christopher

AU - Adams, Peter X.

AU - Amrani, David

AU - Costa, Marco

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AB - Background aims. The distinction between hematopoietic stem cells (HSC) and endothelial progenitor cells (EPC) is poorly defined. Co-expression of CD34 antigen with vascular endothelial growth factor (VEGF) receptor (VEGFR2) is currently used to define EPC (1). Methods. We evaluated the phenotypic and genomic characteristics of peripheral blood-derived CD34+ cells in 22 granulocytecolony-stimulating factor (G-CSF)-mobilized patients with severe coronary artery disease and assessed the influence of cell selection and storage on CD34+ cell characteristics. Results. The median CD34+ cell contents in the products before and after enrichment with the Isolex 300i Magnetic Cell Selection System were 0.2% and 82.5%, respectively. Cell-cycle analysis showed that 80% of CD34+ cells were in G0 stage; 70% of the isolated CD34+ cells co-expressed CD133, a marker for more immature progenitors. However, less than 5% of the isolated CD34+ cells co-expressed the notch receptor Jagged-1 (CD339) and only 2% of the isolated CD34+ population were positive for VEGFR2 (CD309). Molecular assessment of the isolated CD34+ cells demonstrated extremely low expression of VEGFR2 and endothelial nitric oxide synthase (eNOS) and high expression of VEGF-A. Overnight storage at 4°C did not significantly affect CD34+ cell counts and viability. Storage in liquid nitrogen for 7 weeks did not affect the percentage of CD34+ cells but was associated with a 26% drop in cell viability. Conclusions. We have demonstrated that the majority of isolated CD34+ cells consist of immature and quiescent cells that lack prototypic markers of EPC. High VEGF-A gene expression might be one of the mechanisms for CD34+ cell-induced angiogenesis.

KW - Coronary artery disease

KW - Endothelial progenitor cells

KW - Stem cell mobilization

KW - Transplant

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