Monocytes coexpress endothelial and macrophagocytic lineage markers and form cord-like structures in Matrigel® under angiogenic conditions
Monocytes coexpress endothelial and macrophagocytic lineage markers and form cord-like structures in Matrigel® under angiogenic conditions
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DOI：
10.1016/s0008-6363(00)00270-4
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发表时间：
2001-02-16
影响因子：
10.8
通讯作者：
Daniel, WG
中科院分区：
文献类型：
--
作者：
Schmeisser, A;Garlichs, CD;Daniel, WG
目的：研究表明，共表达血管内皮生长因子（VEGF）-R2和AC 133的循环人非贴壁CD 34（+）细胞具有分化为贴壁成熟内皮细胞的能力。然而，先前的研究已经证明，更大的原代贴壁单核细胞亚群也可以形成内皮祖细胞（EPC）。为了确定后一种细胞群体的起源，我们检验了以下假设：单核细胞作为牢固粘附和可塑性细胞类型是否具有分化成内皮表型的潜力。研究方法：采用贴壁分离和磁性弯曲分选法从人外周血中分离CD 34（-）/CD 14（+）单核细胞（纯度>90%），并在纤维连接蛋白包被的塑料培养皿（含VEGF 10 ng/ml、碱性成纤维细胞生长因子（bFGF）2 ng/ml、胰岛素样生长因子（IGF-1）1 ng/ml、20%胎牛血清的培养基）上培养。结果如下：培养2周后，用荧光活化细胞分析法，我们观察到血管内皮标志物vonWillebrand因子（vWf）、VE-cadherin（VE）和ec-NOS分别在45.2%、12.4%和9.8%的细胞中表达。4周后，表达这些标志物的细胞比例进一步增加（分别占这些细胞的94.2%、89.7%和58.8%）。在此期间，表达CD 45的细胞比例保持不变。然而，14天后，特异性巨噬细胞抗原CD 68在62%的分析细胞中新表达，培养28天后进一步增加至90%。在三维凝胶（Matrigel（R））中，观察到索状和管状结构的形成。结论：目前的数据表明，在血管生成的刺激下，巨噬细胞发展的内皮细胞表型与特定的表面标志物的表达，甚至在体外形成索和管样结构，这表明该细胞群可能被招募为血管发生。(C)2001 Elsevier Science B.V，保留所有权利。
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 bend 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(R)) 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. (C) 2001 Elsevier Science B.V, All rights reserved.