Synergistic Neovascularization by Mixed Transplantation of Early Endothelial Progenitor Cells and Late Outgrowth Endothelial Cells

Abstract

Background-Two types of cells are cultured from the human peripheral blood, early endothelial progenitor cells (EPCs) and outgrowth endothelial cells (OECs), as previously reported. Here, we further characterize these cells, especially with respect to their different origins and functions both in vitro and in vivo. We also investigated whether the combination of these different cell types shows synergism during neovascularization. Methods and Results-Early EPCs were heterogeneously made up of both CD14ϩ monocyte-derived cells, which secrete cytokines, and CD14Ϫ -derived cells, which contain high levels of CD34 ϩ KDR ϩ cells. OECs were cultured almost exclusively from CD14 Ϫ cells, not CD14 ϩ cells, and were distinct from mature endothelial cells in terms of proliferation potential, KDR ϩ expression level, and telomerase activity. A portion of cells from CD14 Ϫ cells and early EPCs produced rapidly proliferating, capillary-forming cells in both the Matrigel plug and the ischemic hind limb similar to OECs. Early EPCs and OECs expressed receptors for vascular endothelial growth factor and interleukin-8, cytokines secreted by early EPCs. There was a differential increase in matrix metalloproteinases (MMPs): MMP-9 in early EPCs and MMP-2 in OECs. In vitro, the angiogenic capability of the 2 cell types was augmented by mutual interaction through cytokines and MMPs. Injection of a mixture of the 2 cells resulted in superior neovascularization in vivo to any single-cell-type transplantation. Conclusions-Distinct origins of the different types of EPCs exist that have different functions in neovascularization.Mixed transplantation of these cells results in synergistic neovascularization through cytokines and MMPs.

Endothelial progenitor cells: Implications for cardiovascular disease

“…Since the first report of Asahara et al in 1997, classical isolation methods include adherence culture of total peripheral MNCs (1) and the use of magnetic beads coated with anti-CD34 (1), anti-CD133 (15), or anti-CD14 (16)(17)(18) antibodies. After the isolation, the cells are plated on cell culture dishes coated with either fibronectin (19)(20)(21) or gelatin (8,9,22).…”

Circulating Endothelial Progenitor Cells as a Pathogenetic Marker of Moyamoya Disease

“…Colonies were counted manually in a minimum of 3 wells in 12-well plates by two independent observers who were unaware of clinical profiles; results are expressed as average number of CFUs per well. In selected samples, the endothelial phenotype was confirmed using specific indicators, that is, according to the uptake of 1,1 0 -dioctadecyl-3,3,3 0 ,3 0 -tetramethylindocarbocyanine-labeled acetylated low-density lipoprotein (Biomedical Technologies Inc., Stoughton, MA, USA), and by immunostaining using FITC-labeled Ulex europaeus agglutinin-1 (Sigma) and anti-CD31 (1:20; DAKO, Carpinteria, CA, USA), antivascular endothelium cadherin (1:50; Chemicon, Billerica, MA, USA), anti-CD34 (1:20; DAKO), and anti-kinase domain receptor (1:50; Sigma) antibodies, as previously described (Hur et al, 2004;Yoon et al, 2005;Chu et al, 2008;Jung et al, 2008).…”

“…Culturing PB-MNCs resulted in the formation of CFUs or outgrowth cells, as described previously (Hur et al, 2004;Yoon et al, 2005;Chu et al, 2008;Jung et al, 2008). The interval between blood sampling and cerebral angiography was 15 ± 7 months (range: 3 to 23 months).…”

“…Matters are further complicated by the fact that a subset of outgrowth cells with the endothelial phenotype develops during long-term cultures of peripheral blood mononuclear cells (PBMNCs) (Hur et al, 2004;Yoon et al, 2005;Jung et al, 2008). Outgrowth cells represent an EPC population with vasculogenic potential.…”

Progress and prospects: Cell based regenerative therapy for cardiovascular disease

“…release of various chemokines and cytokines that can affect endogenous cell differentiation, myocardial remodeling and function). [23][24][25][26][27] It has been shown that endothelium-derived growth factors affect the development of apparently unrelated organs and cells, and a paracrine effect has also been demonstrated for other cell types. Thus, Akt-1-transfected mesenchymal progenitor cells have been shown to protect the ischemic heart via a paracrine mechanism.…”