胰岛素对人肝癌HepG2细胞体外诱导人脐静脉内皮细胞血管形成能力的影响

目的:探讨胰岛素对人肝癌细胞系HepG2体外诱导人脐静脉内皮细胞(HUVECs)血管形成能力的影响及其可能机制。方法:用含不同浓度胰岛素的完全培养基预培养肝癌细胞系HepG2制备条件培养基;应用预铺Matrigel基质胶的Transwell小室检测不同组条件培养基对HUVECs迁移能力的影响;运用CCK-8方法及EdU细胞增殖实验检测不同组条件培养基对HUVECs增殖能力的影响;运用内皮细胞成管实验检测不同组条件培养基对HUVECs成血管能力的影响;同时应用RT-PCR检测不同胰岛素浓度培养的HepG2细胞中血管内皮生长因子(VEGF)121、VEGF165、环氧化酶2(COX-2)的转录水平。结果:在一定浓度范围内,HepG2细胞对HUVECs的侵袭迁移能力、增殖能力的影响,对HUVECs的成血管能力的影响,对HepG2细胞VEGF121、VEGF165、COX-2转录水平的影响,均分别与胰岛素浓度呈正相关。结论:在一定浓度范围内,胰岛素可能通过上调HepG2细胞中VEGF121、VEGF165、COX-2的表达水平促进HepG2细胞诱导HUVECs血管形成能力。     

HKD5对人脐静脉内皮细胞游走及血管形成的抑制作用

目的:体外实验研究活化型高分子量激肽原（HKa）轻链富含组氨酸区域5 (HKD5)对人脐静脉内皮细胞（HUVECs）的粘附、游走及血管形成的影响。 方法:体外重组融合蛋白-活化型高分子量激肽原轻链富含组氨酸区域5 (GST-D5H)。WST-1法观察HUVECs细胞粘附能力；用改良Boyden Chamber膜侵袭系统观察HUVECs细胞游走(趋化)；用血管形成实验观察HUVECs细胞形成新生血管能力。 结果:GST-D5H作用后，HUVECs细胞粘贴率降低（P<0.05），游走穿膜细胞数明显低于对照组（P<0.01），诱导内皮细胞形成管腔数及长度均低于对照组（P<0.05）。 结论:GST-D5H能有效抑制HUVECs细胞粘附、游走，使该细胞粘附力降低，迁移性下降及血管形成数目减少或变细。     

肝癌肿瘤微环境对血管内皮细胞增殖及血管生成能力的影响

研究间接共培养条件下肝癌细胞培养基对血管内皮细胞增殖及血管生成能力的影响,初步探讨肿瘤微环境下血管新生的分子机制。体外培养人脐静脉内皮细胞株EA.hy926,与肝癌细胞株HepG2条件培养基进行共培养;四甲基偶氮唑盐(MTT)法检测肝癌细胞条件培养基对血管内皮细胞增殖的影响;血管管腔形成实验检测血管内皮细胞的血管生成能力。Western blot测定肝癌细胞条件培养基对血管内皮EA.hy926细胞血管内皮生长因子(VEGF)及其受体Flk-1表达的影响。结果表明,EA.hy926细胞经HepG2条件培养基处理后,其增殖能力明显增加。接种于Matrigel胶的EA.hy926细胞经HepG2条件培养基刺激后,形成管腔数目增加,成血管能力明显增强;同时,其胞内VEGF及其受体Flk-1的表达呈时间依赖性上调。肿瘤微环境下肝癌细胞作用于血管内皮细胞,可促进血管内皮细胞的增殖并提高其血管生成能力。     

血小板微粒通过激活单核细胞HIF-1α促进血管生成的作用

目的:探讨血小板微粒(PMP)通过单核细胞促进血管生成的作用及机制。方法:制备人PMP,采用基质胶栓实验检测PMP对体内新生血管形成的影响。在体外常氧和缺氧条件下,使PMP与人单核细胞系THP-1结合,ELISA法检测血管内皮生长因子(VEGF)的分泌水平,RT-qPCR检测THP-1细胞VEGF和低氧诱导因子1α(HIF-1α)mRNA的表达,转录活性实验检测THP-1细胞HIF-1α的转录活性;利用共培养系统检测PMP与THP-1细胞相互作用对人脐静脉内皮细胞(HUVECs)体外管腔形成的影响。结果:PMP诱导在体基质胶栓表面形成新生血管,HIF-1α选择性抑制剂chetomin可明显减弱该效应(P0.01)。在常氧和缺氧条件下,PMP均呈剂量依赖性地促进THP-1细胞表达和释放VEGF,上调HIF-1αmRNA的表达并增强其转录活性;阻断PMP与THP-1细胞的相互作用可以抑制THP-1细胞HIF-1α的转录活性以及VEGF的生成。PMP激活的单核细胞可促进HUVECs在体外基质胶上形成管腔,chetomin干预导致管腔的数量明显减少。结论:PMP通过激活单核细胞的HIF-1α诱导其释放VEGF,导致新生血管的形成,这可能是PMP促进动脉粥样硬化血管生成的一种新机制。     

抗人VEGF受体Ⅱ胞外Ⅲ区单克隆抗体的生物学活性研究

目的研究抗KDR单体Ycom1D3抑制VEGF诱导脐静脉内皮细胞（HUVEC）增殖的体外生物学活性。方法采用FACS鉴定Ycom1D3与抗原结合特异性，采用免疫共沉淀测定Ycom1D3阻断VEGF165刺激KDR酪氨酸激酶受体磷酸化作用，并采用[^3H]-Thymidine掺入法、内皮细胞损伤愈合试验和内皮细胞血管腔形成实验进一步确定Ycom1D3抑制VEGF165诱导内皮细胞增殖的中和活性。结果Ycom1D3不但能与HUVEC结合，而且能阻断由VEGF165刺激HUVEC表面KDR酪氨酸激酶受体磷酸化，进而显著抑制VEGF165诱导HUVEC增殖、迁移及体外三维胶原模型上内皮细胞毛细血管样结构的形成。结论Ycom1D3可以通过封闭KDR而抑制VEGF活性，在肿瘤及其它血管新生疾病治疗中具有潜在应用前景。     

VEGF通过上调CCN2促HUVEC迁移和血管生成

目的 探讨血管内皮生长因子(VEGF)诱导的成骨细胞中结缔组织生长因子(CTGF/CCN2)对人脐静脉血管内皮细胞(HUVECs)的影响.方法 用Real time PCR法及ELISA法检测VEGF诱导成骨细胞(OSE)中CCN2含量;制备成骨细胞(OSE)上清液;将细胞分为control组、OSE组和VEGF-OSE组(n=3).用小干扰RNA (siRNA)转染法抑制成骨细胞中CCN2的表达;Transwell法检测内皮细胞迁移;Matrigel实验检测管样结构形成能力.结果 VEGF呈时间和剂量依赖性上调成骨细胞中CCN2 mRNA和蛋白的表达;CCN2可促进内皮细胞的迁移和管样结构形成(P＜0.05),当CCN2被siRNA基因沉默或者加入CCN2抗体后,CCN2对内皮细胞迁移和管样结构形成的促进作用均受到明显抑制(P＜0.05).结论 VEGF通过上调成骨细胞中CCN2的表达,促内皮细胞(HUVECs)的迁移和血管生成.     

CD133+卵巢癌干细胞样细胞在血管内皮细胞中的分化

背景：大量研究证实,新生血管形成在肿瘤的生长、浸润以及转移过程中发挥重要作用。 目的：探讨CD133+卵巢癌干细胞样细胞向血管内皮细胞分化的特点。 方法：通过无血清培养方法从卵巢癌A2780细胞株中成功诱导出CD133⁺卵巢癌干细胞样细胞,在体外接种于铺或不铺Matrigel基质胶的96孔板内,观察不同时间点CD133⁺卵巢癌干细胞样细胞和人脐静脉内皮细胞形成管腔样结构能力。通过裸鼠皮下移植实验,免疫荧光法观察CD133⁺卵巢癌干细胞样细胞在卵巢癌血管新生中的作用。 结果与结论：CD133⁺卵巢癌干细胞样细胞和人脐静脉内皮细胞(阳性对照)在未铺 Matrigel基质胶上并不能形成相应的管腔结构,且不表达内皮细胞标志物CD31,在Matrigel基质胶上能够形成相对稳定的管腔结构,CD31表达明显。CD133⁺卵巢癌干细胞样细胞接种裸鼠皮下成瘤后,可观察到肿瘤组织中有人源性CD31的表达。结果表明CD133⁺卵巢癌干细胞样细胞能够分化为血管内皮细胞,参与肿瘤血管重建。中国组织工程研究杂志出版内容重点：干细胞;骨髓干细胞;造血干细胞;脂肪干细胞;肿瘤干细胞;胚胎干细胞;脐带脐血干细胞;干细胞诱导;干细胞分化;组织工程     

转录因子Bach1对人微血管内皮细胞的作用研究

 目的:研究转录因子Bach1对人微血管内皮细胞功能的影响。方法: 利用小干扰RNA（small interfering RNA,siRNA）细胞转染技术下调内皮细胞Bach1表达;用Matrigel管腔形成实验检测内皮细胞体外血管新生的能力;用Transwell小室法检测细胞迁移;用CCK-8法测定细胞增殖;用实时荧光定量PCR、Western blotting和ELISA法检测细胞中血红素氧合酶1（heme oxygenase 1,HO-1）和血管内皮细胞生长因子（vascular endothelial growth factor, VEGF）mRNA 和蛋白的表达情况;用转染报告基因的方法检测VEGF基因的转录活性。结果: 下调内皮细胞Bach1表达明显促进人微血管内皮细胞迁移和管腔形成能力,对内皮细胞增殖能力无明显影响;抑制Bach1表达促进内皮细胞HO-1 mRNA 和蛋白的表达,增加VEGF 转录活性及mRNA和蛋白的表达。结论: 抑制转录因子Bach1表达可增加内皮细胞HO-1和VEGF的表达,促进人微血管内皮细胞迁移和管腔形成,提示Bach1是负性调控血管新生的因子。     

血管内皮生长因子诱导的胶质瘤源性微血管内皮细胞体外三维成型特性

目的比较人脑胶质瘤源性微血管内皮细胞（GDMEC）和内皮细胞样细胞ECV304三维培养血管生成特性,探讨GDMEC在血管生成研究中的意义。方法采用免疫磁珠分选系统获得纯化的人脑GDMEC,以胶原为介质建立内皮细胞三维培养模型,比较观察GDMEC与ECV304细胞三维培养小管样结构（TLS）形成及不同浓度血管内皮生长因子（VEGF）对两种细胞TLS形成的诱导作用。结果所获GDMEC细胞纯度达98％,可连续传代培养。GDMEC的TLS形成数显著多于同数量、同时相点ECV304细胞的TLS形成数量。VEGF在体外对GDMEC有显著的促进TLS形成作用,且呈量-效和时-效关系;VEGF对ECV304细胞形成小管样结构的诱导作用弱。结论GDMEC在体外保持了内皮细胞特征和活跃的血管生成特性,对VEGF的反应性较ECV304好,因而GDMEC更适合体外血管生成研究。     

Transwell侵袭小室技术的改良及其在人骨髓间充质干细胞诱导分化中的应用

目的：分离、培养和鉴定人骨髓间充质干细胞（Mesenchymal stem cells，MSCs），应用改良的Transwell侵袭小室技术，探讨血管内皮生长因子（VEGF）及内皮细胞条件诱导液对其体外诱导分化中的作用。方法：采用Percoll（1．073g／ml）分离液分离骨髓单个核细胞，体外培养MSCs，流式细胞术分析鉴定MSCs的纯度，Transwell侵袭小室技术结合LSCM，实时监测MSCs在Matrigel与VEGF／内皮细胞条件诱导液构成的内皮细胞生长微环境中的运动迁移情况。结果：经Percoll分离、体外培养扩增的MSCs，细胞纯度可达95％左右；VEGF组迁移的深度虽高于对照组（P〈0．05），但迁移至聚碳酸脂膜下的细胞与对照组相比，并无统计学意义（P〉0．05）。内皮细胞条件诱导液促进MSCs的迁移，在Matrigel内迁移的深度及迁移至聚碳酸脂膜下的细胞均明显多于对照组（P〈0．05）。结论：共聚焦激光扫描显微术与Transwell侵袭小室技术的结合，能够从时间和空间上对后者进行观察，使该实验得到改良；利用内皮细胞条件诱导液与Matrigel模拟体外内皮细胞生长的微环境，并从空间上观测了MSCs穿越人工基底膜的情况，为MSCs向内皮细胞体外诱导开辟了新的思路。     

GM-CSF induces expression of soluble VEGF receptor-1 from human monocytes and inhibits angiogenesis in mice

GM-CSF promotes homeostasis of myeloid cells. We report that GM-CSF upregulates mRNA and protein production of the soluble form of membrane bound VEGF receptor-1 (sVEGFR-1) in human monocytes. This sVEGFR-1 was biologically active, as cell-free supernatants from GM-CSF-stimulated monocytes blocked detection of endogenously expressed VEGF and inhibited endothelial cell migration and tube formation, even in the presence of exogenous rhVEGF. VEGF activity was recovered by neutralizing sVEGFR-1. To determine whether these events were important in vivo, Matrigel plugs were incubated with rhVEGF, rhGM-CSF, or rhGM-CSF/rhVEGF and injected into mice. Plugs containing GM-CSF or GM-CSF/VEGF had less endothelial cell invasion than plugs containing rhVEGF and were similar to plugs incubated with PBS alone. Neutralizing antibodies specific for sVEGFR-1 injected in these plugs reversed the effects of GM-CSF or GM-CSF/VEGF, while an isogenic antibody did not. Thus, GM-CSF and monocytes play a vital role in angiogenesis through the regulation of VEGF and sVEGFR-1.     

VEGF基因转染脂肪干细胞后上清液对皮肤成纤维细胞及脐静脉血管内皮细胞的影响

目的:通过慢病毒将血管内皮生长因子(VEGF)基因转染于人脂肪干细胞(HADSCs),检测其上清液(CM)中生长因子的表达,并用其上清液作用于人皮肤成纤维细胞(HDFs)及人脐静脉内皮细胞(HUVECs),观察对这2种细胞活力和迁移的影响。方法:准备HADSCs、HDFs及HUVECs 3种细胞并鉴定;将携带VEGF165基因的慢病毒转染于HADSCs,定时收集上清液;ELISA法检验上清中生长因子分泌情况;将VEGF-CM与完全培养液以一定比例混合分为5组,分别培养HDFs及HUVECs,CCK-8法检验对2种细胞活力的影响;将最佳比例的VEGF-CM、正常CM(Nor-CM)和完全培养液分别作用于HDFs及HUVECs,划痕法测试出对2种细胞迁移的影响。结果:ELISA结果表明VEGF-CM中VEGF及碱性成纤维细胞生长因子(bFGF)的表达均较Nor-CM组提高;相对于其它比例,当完全培养液与VEGF-CM以1∶2的比例混合时,HDFs和HUVECs的活力显著增强(P0.05);VEGF-CM与其它培养液相比可显著提高HDFs和HUVECs的迁移能力(P0.05)。结论:转染VEGF165基因后的HADSCs可同时增强VEGF及bFGF的分泌,其上清液可提高成纤维细胞及血管内皮细胞的活力和迁移能力。     

Bovine lactoferricin inhibits basic fibroblast growth factor- and vascular endothelial growth factor165-induced angiogenesis by competing for heparin-like binding sites on endothelial cells

Angiogenesis is a complex process whereby new blood vessels form from pre-existing vasculature in response to proangiogenic factors such as basic fibroblast growth factor (bFGF) and the 165-kd isoform of vascular endothelial growth factor (VEGF165). Angiogenesis inhibitors show considerable potential in the treatment of cancer because angiogenesis is necessary for tumor growth beyond a few millimeters in diameter because of the tumor's need for oxygen and nutrient supply, as well as waste removal. Bovine lactoferricin (LfcinB) is a peptide fragment of iron- and heparin-binding lactoferrin obtained from cow's milk. Here we provide in vivo and in vitro evidence that LfcinB has potent antiangiogenic activity. LfcinB strongly inhibited both bFGF- and VEGF165-induced angiogenesis in Matrigel plugs implanted in C57BL/6 mice. In addition, LfcinB inhibited the in vitro proliferation and migration of human umbilical vein endothelial cells (HUVECs) in response to bFGF or VEGF165 but was not cytotoxic to HUVECs. Rather, LfcinB complexed with heparin-like structures on the HUVEC surface that are involved in the binding of bFGF and VEGF165 to their respective receptors, thereby preventing receptor-stimulated angiogenesis. These findings suggest that LfcinB may have utility as an antiangiogenic agent for the treatment of human cancers.     

Vascular Endothelial Growth Factor Receptor-1 Modulates Vascular Endothelial Growth Factor-Mediated Angiogenesis via Nitric Oxide

The known responses of vascular endothelial growth factor (VEGF) are mediated through VEGF receptor-2 (VEGFR-2/KDR) in endothelial cells. However, it is unknown whether VEGFR-1 (Flt-1) is an inert decoy or a signaling receptor for VEGF during physiological or pathological angiogenesis. Here we report that VEGF-stimulated nitric oxide (NO) release is inhibited by blockade of VEGFR-1 and that VEGFR-1 via NO negatively regulates of VEGFR-2-mediated proliferation and promotes formation of capillary networks in human umbilical vein endothelial cells (HUVECs). Inhibition of VEGFR-1 in a murine Matrigel angiogenesis assay induced large aneurysm-like structures. VEGF-induced capillary growth over 14 days was inhibited by anti-VEGFR-2-blocking antibody as determined by reduced tube length between capillary connections (P < 0.0001) in an in vitro angiogenesis assay. In contrast, loss of VEGFR-1 activity with a neutralizing anti-VEGFR-1 antibody resulted in an increase in the accumulation of endothelial cells (P < 0.0001) and a dramatic decrease in the number of capillary connections that were restored by the addition of NO donor. Porcine aortic endothelial (PAE) cells expressing human VEGFR-1 but not VEGFR-2 plated on growth factor-reduced Matrigel rearranged into tube-like structures that were prevented by anti-VEGFR-1 antibody or a cGMP inhibitor. VEGF stimulated NO release from VEGFR-1- but not VEGFR-2-transfected endothelial cells and placenta growth factor-1 stimulated NO release in HUVECs. Blockade of VEGFR-1 increased VEGF-mediated HUVEC proliferation that was inhibited by NO donors, and potentiated by NO synthase inhibitors. These data indicate that VEGFR-1 is a signaling receptor that promotes endothelial cell differentiation into vascular tubes, in part by limiting VEGFR-2-mediated endothelial cell proliferation via NO, which seems to be a molecular switch for endothelial cell differentiation.     

B7-H3 modulates endothelial cell angiogenesis through the VEGF cytokine

B7-H3 is a cell surface molecule in the immunoglobulin superfamily that has been shown to perform both immunological and non-immunological functions. It has also been found that vascular endothelial growth factor (VEGF) is an important molecule in the modulation of endothelial cell behavior. In this study, we analyzed the serum expression of B7-H3 in 113 rheumatoid arthritis and systemic lupus erythematous patients using the ELISA and found a positive correlation between B7-H3 and VEGF. Next, we investigated the involvement of B7-H3 in angiogenesis using human umbilical vein endothelial cells (HUVECs) with transient knockdown of B7-H3 and an in vivo Matrigel model. Data from the in vitro experiments showed that B7-H3 increased cell proliferation, migration, and tube formation, and correlated with the expression of VEGF. Furthermore, B7-H3 affected the formation of functional vascular networks in Matrigel plugs, which were dissected from mice injected with different HUVECs. Our data suggest that B7-H3 promotes angiogenesis through the enhancement of VEGF secretion. This is the first study proposing a significant role for B7-H3 in the promotion of angiogenesis and may provide further understanding of this gene’s biological function.

     

Distinct patterns of microvascular endothelial cell morphology are determined by extracellular matrix composition.

Endothelial interactions with the extracellular matrix (ECM) play important roles in angiogenesis but whether specific ECM signals can determine specific cellular morphologies is unclear. The authors compared in vitro ECM-induced morphological responses of the phenotypically distinct human placental microvascular endothelial cells (HPMECs) with large vessel endothelial cells (HUVECs). HPMECs showed distinct patterns of reorganization in response to collagen-I or collagen-IV (monolayer disruption, sprouting, migration) and Matrigel or laminin-A (intussusception, cord formation, tubulogenesis), and an intermediate response to fibrin; whereas HUVECs responded similarly to collagen-1 and Matrigel (elongation, lattice formation, vacuolation) and showed little response to fibrin. Although the extent of collagen and Matrigel responses of HPMECs were increased by serum, acidic or basic fibroblast growth factor (aFGF, bFGF), or vascular endothelial growth factor (VEGF), and varied with matrix protein concentration, the basic patterns were matrix specific, and were independent of fibronectin. The collagen responses correlated with disruption of adherens and tight junctions and the formation of filopodial protrusions. Matrigel responses were associated with up-regulated junctional localization of VE-cadherin, and tubulogenesis developed mainly through paracellular remodeling rather than intracellular vacuolation. Overall, these findings suggest that distinct ECM interactions stimulate specific morphological responses. These signals may regulate morphological behaviour in the angiogenesis cycle, switching endothelial cells between migratory and vasculogenic phenotypes.     

丁苯酞通过激活VEGF/VEGFR2-Notch1/Dll4信号促进HUVECs形成血管

目的:探索缺血缺氧(H/I)条件下丁苯酞(NBP)通过激活血管内皮生长因子(VEGF)/VEGF受体2(VEGFR2)-Notch1/Delta样配体4(Dll4)信号促进人脐静脉内皮细胞(HUVECs)血管形成的机制。方法:利用无血清培养基和缺氧罐模拟H/I条件,HUVECs传代培养后设置正常对照(control)组、H/I组、NBP高剂量(H/I+NBP_(high))组和NBP低剂量(H/I+NBP_(low))组,其中control组为常规培养的HUVECs,H/I组为H/I条件下培养的HUVECs,H/I+NBP_(high)组为在H/I环境下使用20μmol/L丁苯酞干预的HUVECs,H/I+NBP_(low)组为在H/I环境下使用5μmol/L丁苯酞干预的HUVECs。CCK-8法检测各组细胞的细胞活力,细胞划痕实验检测各组细胞的迁移能力,体外血管形成实验检测各组细胞成管能力,Western blot法检测VEGFR2、Notch1和Dll4的表达,ELISA法检测培养基中VEGF的表达,qPCR检测VEGF、VEGFR2、Notch1和Dll4的mRNA表达水平。结果:丁苯酞可以提高H/I条件下HUVECs的存活率,促进细胞的迁移和体外血管的形成能力,且H/I+NBP_(high)组比H/I+NBP_(low)组更加显著。丁苯酞可以提高VEGF、VEGFR2、Notch1和Dll4的mRNA及蛋白表达。结论:丁苯酞可以在H/I条件下促进HUVECs形成血管,其机制可能与VEGF/VEGFR2-Notch1/Dll4信号的激活有关。     

口虾蛄提取物对人鼻咽癌细胞迁移和体外血管生成拟态的抑制作用

 目的：研究口虾蛄提取物（extract of Oratosquilla,EOS）对人低分化鼻咽癌细胞株CNE-2的迁移及体外血管生成拟态的影响。方法：用不同浓度（0 mg/L 、125 mg/L、250 mg/L、500 mg/L）EOS处理CNE-2细胞24 h后,创伤修复实验检测细胞迁移能力;通过Matrigel三维细胞培养观察CNE-2细胞形成类血管网状结构的能力及其特点;体外管道形成抑制实验检测不同浓度EOS对CNE-2细胞管道形成能力的影响;Western blotting法检测不同浓度EOS对CNE-2细胞fascin 1和血管内皮生长因子（VEGF）蛋白表达的影响。结果：EOS可以显著降低CNE-2细胞的迁移能力,与对照组比较差异有统计学意义（P＜0.01）;CNE-2细胞在Matrigel上培养能形成类似血管的网状样结构;EOS能以剂量依赖方式抑制CNE-2细胞体外管道形成的数量（P＜0.01）;EOS能抑制CNE-2细胞中fascin 1和VEGF蛋白的表达（P＜0.01）,且其管状结构数量与2种蛋白变化趋势呈正相关（P＜0.05）。结论：CNE-2细胞具有血管生成拟态的能力;EOS能够抑制CNE-2细胞的迁移和体外血管生成拟态的能力,其机制可能与下调fascin 1和VEGF蛋白的表达有关。     

内皮祖细胞在体外培养成血管样结构的初步观察

目的：探索体外培养脐血、外周血内皮祖细胞（EPCs）的方法，观察其形成血管样结构的可能性及条件。 方法： 采用贴壁选择法培养人脐血及兔外周血内皮祖细胞，光镜下观察细胞形态，用荧光显微镜、流式细胞仪分析贴壁细胞CD34、VEGFR-2、AC133、血管内皮钙粘素（VE-cadherin）的表达，DiI-ac-LDL 吞噬试验及Ⅷ因子免疫组化证实细胞属性。 结果： 体外成功培养出人脐血及兔外周血内皮祖细胞，形成条索状、管状结构，兔外周血EPCs分化较成熟，形成典型铺路石形状及血管样结构。 结论： 脐血、兔外周血内皮祖细胞可在体外培养成功并表现成血管倾向，可能是血管组织工程的潜在资源。