Angiogenic potential of early and late outgrowth endothelial progenitor cells is dependent on the time of emergence
Angiogenic potential of early and late outgrowth endothelial progenitor cells is dependent on the time of emergence
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DOI：
10.1016/j.ijcard.2015.03.166
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发表时间：
2015-05-01
影响因子：
3.5
通讯作者：
Sahara, Makoto
中科院分区：
文献类型：
--
作者：
Minami, Yoshiyasu;Nakajima, Toshiaki;Sahara, Makoto
背景：最近的研究表明，人外周血单个核细胞(HPBMNCs)来源的晚期生长内皮祖细胞(EPC)可能比经典定义的早期生长内皮祖细胞(EOCs)具有更高的血管生成潜力。然而，目前还不清楚以不同方式定义的所谓的EPC亚群中哪些具有最高的血管生成潜力。方法和结果：我们根据hPBMNC来源的EPC亚群在培养中出现的时间对其进行分类。EOCs定义为培养3-7天的贴壁细胞。晚期生长内皮祖细胞是指从第10天开始形成鹅卵石状集落的细胞，其分类如下：“中等”--第10-16天出现的生长内皮祖细胞(MOCs)，“晚期”--第17-23天出现的生长内皮祖细胞(LOC)，以及“非常晚”--第24-30天出现的生长内皮祖细胞(VOCs)。流式细胞仪分析显示EOC(CD31(+)VE-cadherin-CD34(-)CD14(+)CD45(+))与LOC(CD31(+)VE-cadherin(+)CD34(+)CD14(-)CD45(-))的造血/内皮标志物有明显差异。我们发现，LOC在体外具有最高的增殖和管状形成能力，并具有最高的血管生成基因表达，包括KDR和eNOS。为探讨EPC亚群的体内治疗效果，将各EPC亚群静脉移植到免疫功能低下的小鼠(共4×10(5)个细胞)中。LOC组小鼠缺血小腿血流恢复显著增强(缺血侧/非缺血侧血流比率：0.99+/-0.02[LOC组]：0.67+/-0.07比0.78+/-0.09[其他组]；P<；0.05)，缺血小腿毛细血管侧支形成增加，这归因于其直接植入宿主血管生成血管(约10%)和旁分泌效应。结论：培养17-23天的hPBMNC来源的晚期生长EPC亚群具有更好的治疗血管生成潜力。(C)2015爱思唯尔爱尔兰有限公司。保留所有权利。
Background: Recent studies have suggested that late-outgrowth endothelial progenitor cells (EPCs) derived from human peripheral blood mononuclear cells (hPBMNCs) might have higher angiogenic potential than classically-defined early-outgrowth EPCs (EOCs). However, it still remains unclear which of "so-called" EPC subpopulations defined in a variety of ways has the highest angiogenic potential.Methods and results: We classified hPBMNC-derived EPC subpopulations by the time of their emergence in culture. EOCs were defined as attached cells on culture days 3-7. Late-outgrowth EPCs, defined as the cell forming colonies with cobblestone appearance since day 10, were further classified as follows: "moderate"-outgrowth EPCs (MOCs) emerging on days 10-16, "late"-outgrowth EPCs (LOCs) on days 17-23, and "very late"-outgrowth EPCs (VOCs) on days 24-30. Flow cytometry analyses showed the clear differences of hematopoietic/endothelial markers between EOC (CD31(+)VE-cadherin-CD34(-)CD14(+)CD45(+))and LOC (CD31(+)VE-cadherin(+)CD34(+)CD14(-)CD45(-)). We found that LOCs had the highest proliferation and tube formation capabilities in vitro along with the highest expression of angiogenic genes including KDR and eNOS. To investigate the in vivo therapeutic efficacies, each EPC subpopulation was intravenously transplanted into immunocompromised mice (total 4 x 10(5) cells) after unilateral hindlimb ischemia surgery. The LOC-treatedmice exhibited significantly-enhanced blood flow recovery (flow ratios of ischemic/non-ischemic leg: 0.99 +/- 0.02 [LOC group] versus 0.67 +/- 0.07 to 0.78 +/- 0.09 [other groups]; P < 0.05) and augmented capillary collateral formation in ischemic leg, which were attributable to their direct engraftment into host angiogenic vessels (approximately 10%) and paracrine effects.Conclusion: hPBMNC-derived late-outgrowth EPCs emerging on culture days 17-23 are superior to other EPC subpopulations with regard to therapeutic angiogenic potential. (C) 2015 Elsevier Ireland Ltd. All rights reserved.