Finney Marcie R, Greco Nicholas J, Haynesworth Stephen E, Martin Joseph M, Hedrick David P, Swan Jimmy Z, Winter Daniel G, Kadereit Suzanne, Joseph Matthew E, Fu Pingfu, Pompili Vincent J, Laughlin Mary J
Department of Medicine, Case Western Reserve University, School of Medicine, Case Comprehensive Cancer Center, Cleveland, Ohio 44106-7284, USA.
Biol Blood Marrow Transplant. 2006 May;12(5):585-93. doi: 10.1016/j.bbmt.2005.12.037.
Endothelial precursor cells (EPCs) cultured from adult bone marrow (BM) have been shown to mediate neovasculogenesis in murine models of vascular injury. We sought to directly compare umbilical cord blood (UCB)- and BM-derived EPC surface phenotypes and in vivo functional capacity. UCB and BM EPCs derived from mononuclear cells (MNC) were phenotyped by surface staining for expression of stromal (Stro-1, CXCR4, CD105, and CD73), endothelial (CD31, CD146, and vascular endothelial [VE]-cadherin), stem cell (CD34 and CD133), and monocyte (CD14) surface markers and analyzed by flow cytometry. The nonobese diabetic/severe combined immunodeficiency murine model of hind-limb ischemia was used to analyze the potential of MNCs and culture-derived EPCs from UCB and BM to mediate neovasculogenesis. Histologic evaluation of the in vivo studies included capillary density as a measure of neovascularization. Surface CXCR4 expression was notably higher on UCB-derived EPCs (64.29%+/-7.41%) compared with BM (19.69%+/-5.49%; P=.021). Although the 2 sources of EPCs were comparable in expression of endothelial and monocyte markers, BM-derived EPCs contained higher proportions of cells expressing stromal cell markers (CD105 and CD73). Injection of UCB- or BM-derived EPCs resulted in significantly improved perfusion as measured by laser Doppler imaging at days 7 and 14 after femoral artery ligation in nonobese diabetic/severe combined immunodeficiency mice compared with controls (P<.05). Injection of uncultured MNCs from BM or UCB showed no significant difference from control mice (P=.119; P=.177). Tissue samples harvested from the lower calf muscle at day 28 demonstrated increased capillary densities in mice receiving BM- or UCB-derived EPCs. In conclusion, we found that UCB and BM-derived EPCs differ in CXCR4 expression and stromal surface markers but mediate equivalent neovasculogenesis in vivo as measured by Doppler flow and histologic analyses.
已证明，从成年骨髓（BM）培养的内皮祖细胞（EPC）在血管损伤的小鼠模型中介导新生血管形成。我们试图直接比较脐带血（UCB）和BM来源的EPC表面表型及体内功能能力。通过对单核细胞（MNC）来源的UCB和BM EPC进行表面染色，检测基质（Stro-1、CXCR4、CD105和CD73）、内皮（CD31、CD146和血管内皮[VE] -钙黏蛋白）、干细胞（CD34和CD133）以及单核细胞（CD14）表面标志物的表达，进行表型分析，并通过流式细胞术进行检测。采用后肢缺血的非肥胖糖尿病/严重联合免疫缺陷小鼠模型，分析来自UCB和BM的MNC及培养的EPC介导新生血管形成的潜力。体内研究的组织学评估包括将毛细血管密度作为新生血管形成的指标。与BM来源的EPC（19.69%±5.49%）相比，UCB来源的EPC表面CXCR4表达显著更高（64.29%±7.41%；P = 0.021）。尽管这两种EPC来源在内皮和单核细胞标志物的表达上相当，但BM来源的EPC中表达基质细胞标志物（CD105和CD73）的细胞比例更高。在非肥胖糖尿病/严重联合免疫缺陷小鼠中，与对照组相比，在股动脉结扎后第7天和第14天，通过激光多普勒成像测量发现，注射UCB或BM来源的EPC可显著改善灌注（P < 0.05）。注射来自BM或UCB的未培养MNC与对照小鼠相比无显著差异（P = 0.119；P = 0.177）。在第28天从小腿下部肌肉采集的组织样本显示，接受BM或UCB来源EPC的小鼠毛细血管密度增加。总之，我们发现UCB和BM来源的EPC在CXCR4表达和基质表面标志物方面存在差异，但通过多普勒血流和组织学分析测量，它们在体内介导的新生血管形成相当。
