© 2001 by European Society of Cardiology
Copyright © 2001, European Society of Cardiology
Monocytes coexpress endothelial and macrophagocytic lineage markers and form cord-like structures in Matrigel® under angiogenic conditions
aDepartment of Cardiology, Technical University of Dresden, Heart Center Dresden, Fetscherstr. 76, D-01307 Dresden, Germany
bDivision of Molecular Cardiology, Department of Cardiology, University Erlangen-Nuernberg, Oestliche Stadtmauerstr. 29, D-91054 Erlangen, Germany
* Corresponding author. Tel.: +49-351-450-1704; fax: +49-351-450-1702 alexanderschmeis{at}t-online.de
Objectives: It has been shown that circulating human non-adherent CD34+ cells coexpressing vascular endothelial growth factor (VEGF)-R2 and AC133 have the capacity to differentiate into adherent mature endothelial cells. However, prior studies have demonstrated that a much bigger subset of primary adherent mononuclear cells can also form endothelial progenitor cells (EPC). To determine the origin of the latter cell population we tested the hypothesis: do monocytes as a firmly adherent and plastic cell type have the potential to differentiate into an endothelial phenotype. Methods: CD34–/CD14+ monocytes were isolated from human peripheral blood by adherence separation and magnetic bead selection (purity <90%) and cultured on fibronectin-coated plastic dishes (medium containing VEGF 10 ng/ml, basic fibroblast growth factor (bFGF) 2 ng/ml, insulin like growth factor (IGF-1) 1 ng/ml, 20% fetal calf serum). Results: After 2 weeks of culture, using fluorescence activated cell analysis we observed a new expression of the endothelial markers von Willebrand factor (vWf), VE-cadherin (VE) and ec-NOS in 45.2, 12.4 and 9.8% of the cells, respectively. The proportion of cells expressing these markers further increased after 4 weeks (94.2, 89.7 and 58.8% of these cells, respectively). The proportion of CD45 expressing cells remained unchanged during this period. However, after 14 days the specific macrophage antigen CD68 was newly expressed in 62% of the analysed cells with a further increase to 90% after 28 days of culture. In three-dimensional gel (Matrigel®) the formation of cord- and tubular-like structures was observed. Conclusion: The present data indicate that under angiogenic stimulation macrophages develop an endothelial phenotype with expression of specific surface markers and even form cord- and tubular-like structures in vitro suggesting that this cell population may be recruited for vasculogenesis.
KEYWORDS bFGF, basic fibroblast growth factor; EC, endothelial cells; EPC, endothelial progenitor cells; FACS, fluorescence activated cell analysis; FCS, fetal calf serum; Flk-1, fetal liver kinase-1; Flt-1, fms-like tyrosine kinase-1; HSC, hematopoietic stem cells; HUVEC, human umbilical vein endothelial cells; IGF-1, insulin like growth factor; Mo, monocytes; ox-LDL, oxidized low density lipoprotein; VE, VE-cadherin; VEGF, vascular endothelial growth factor; vWf, von Willebrand factor
1 These authors have contributed equally to this work.
* Presented in part at the 72nd Scientific Session of the American Heart Association, Atlanta, GA, November 7–10, 1999, and published in abstract form [Circulation 100 (1999) I-405]. During the review process of this manuscript similar findings where published by Fernandez Pujol et al. [Differentiation 65 (2000) 287–300].
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