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Author Notes:

Address for Correspondence: Kreton Mavromatis, MD 1670 Clairmont Rd, 111B Decatur, GA 30033 Tel: 404-329-2207; Fax: 404-329-2211 kmavro@emory.edu

The authors declared no potential conflicts of interest with respect to the research, authorship, and/or publication of this article.

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Research Funding:

The authors disclosed receipt of the following financial support for the research, authorship, and/or publication of this article: This work was supported by Eli Lilly and Company, Indianapolis, Indiana; a gift from the Marcus Foundation, Atlanta, Georgia; the Georgia Tech/Emory Center for the Engineering of Living Tissues (GTEC) National Science Foundation (NSF) Grant EEC-9731643; PHS Grant UL1 RR025008 from the Clinical and Translational Science Award program, National Institutes of Health, National Center for Research Resources; and the Woodruff Fund.

Keywords:

  • stem cells
  • cardiac disease
  • cell-based assays

Pro-angiogenic Cell Colonies Grown In Vitro from Human Peripheral Blood Mononuclear Cells

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Journal Title:

Journal of Biomolecular Screening

Volume:

Volume 17, Number 9

Publisher:

, Pages 1128-1135

Type of Work:

Article | Post-print: After Peer Review

Abstract:

Although multiple culture assays have been designed to identify “endothelial progenitor cells” (EPCs), the phenotype of cells grown in culture often remains undefined. We sought to define and characterize the pro-angiogenic cell population within human peripheral blood mononuclear cells. Mononuclear cells were isolated from peripheral blood and grown under angiogenic conditions for 7 days. Formed colonies (CFU-As) were identified and analyzed for proliferation, mRNA and surface antigen expression, tube-forming ability and chromosomal content. Colonies were composed of a heterogeneous group of cells expressing the leukocyte antigens CD45, CD14, and CD3, as well as the endothelial proteins vascular endothelial (VE) cadherin, von Willebrand's Factor (vWF), CD31 and endothelial nitric oxide synthase (eNOS). Colony cells expressed increased levels of pro-angiogenic growth factors, and they formed tubes in Matrigel. In comparison with colonies from the CFU-Hill assay, our assay resulted in a greater number of colonies (19±9 vs. 13±7; p<0.0001) with a substantial number of cells expressing an endothelial phenotype (20.2±7.4% vs. 2.2±1.2% expressing eNOS, p=0006). Chromosomal analysis indicated the colony cells were bone marrow-derived. We, therefore, describe a colony forming unit assay that measures bone marrow-derived circulating mononuclear cells with the capacity to proliferate and mature into proangiogenic leukocytic and endothelial-like cells. This assay, therefore, reflects circulating, bone marrow-derived pro-angiogenic activity.

Copyright information:

© 2012, Society for Laboratory Automation and Screening

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