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脐带血中筛选出的 CD133(+) 细胞在体外和体内均表现出血管生成功能。

Umbilical cord blood-selected CD133(+) cells exhibit vasculogenic functionality in vitro and in vivo.

机构信息

Department of Medicine, Case Western Reserve University, School of Medicine, Cleveland, Ohio, USA.

出版信息

Cytotherapy. 2010;12(1):67-78. doi: 10.3109/14653240903300658.

Abstract

BACKGROUND AIMS

Current clinical trials utilize non-selected bone marrow (BM) mononuclear cells (MNC) to augment vasculo genesis within ischemic vascular beds. Recent reports have identified a diminished number and function of hemat-opoietic stem cells (HSC) from aged and diseased patients. Umbilical cord blood (UCB) provides a potential robust allo-geneic source of HSC for therapeutic vasculogenesis.

METHODS

MNC and magnetically isolated CD133(+) cells were assessed for viability (trypan blue) and surface phenotype (flow cytometry). To test in vivo functionality of the cells, NOD/SCID mice underwent ligation of the right femoral artery followed immediately by cell injection. Blood flow recovery, necrosis, BM engraftment of human cells and histologic capillary density were determined. Cells were tested for potential mechanisms mediating the in vivo effects, including migration, cytokine secretion and angiogenic augmentation (Matrigel assays).

RESULTS

Surface expression analysis showed CD31 (PECAM) expression was greatly increased in UCB CD133(+) cells compared with BM MNC. At 28 days, perfusion ratios were highest in animals receiving UCB CD133(+) cells, while animals receiving BM CD133(+) cells and BM MNC demonstrated perfusion ratios statistically higher than in animals treated with cytokine media alone. Animals receiving CD133(+) cells showed a statistically higher capillary density, reduced severe digit necrosis and increased engraftment in the BM than animals treated with unselected BM MNC. In vitro studies showed equivalent migration to stromal-derived factor-1 (SDF-1), increased production of tumor necrosis factor alpha (TNF-alpha) and increased branch points with the co-incubation of CD133(+) cells with human umbilical vein endothelial cells (HUVEC) in the Matrigel angiogenesis assay.

CONCLUSIONS

Taken together, UCB CD133(+) cells exhibit robust vasculogenic functionality compared with BM MNC in response to ischemia.

摘要

背景目的

目前的临床试验利用非选择性骨髓(BM)单核细胞(MNC)来增加缺血性血管床内的血管生成。最近的报告表明,来自老年和患病患者的造血干细胞(HSC)数量和功能减少。脐带血(UCB)为治疗性血管生成提供了潜在的强大同种异体 HSC 来源。

方法

评估 MNC 和磁性分离的 CD133(+)细胞的活力(台盼蓝)和表面表型(流式细胞术)。为了测试细胞的体内功能,NOD/SCID 小鼠接受右侧股动脉结扎,随后立即注射细胞。通过血流恢复、坏死、BM 中人类细胞的植入和组织学毛细血管密度来确定。测试了细胞潜在的介导体内效应的机制,包括迁移、细胞因子分泌和血管生成增强(Matrigel 测定)。

结果

表面表达分析显示,与 BM MNC 相比,UCB CD133(+)细胞的 CD31(PECAM)表达大大增加。在 28 天,接受 UCB CD133(+)细胞的动物的灌注比最高,而接受 BM CD133(+)细胞和 BM MNC 的动物的灌注比与仅接受细胞因子介质治疗的动物相比统计学上更高。接受 CD133(+)细胞的动物的毛细血管密度较高,严重的指端坏死减少,并且在 BM 中的植入物比接受未选择的 BM MNC 的动物高。体外研究表明,在基质衍生因子-1(SDF-1)的趋化作用、肿瘤坏死因子-α(TNF-α)的产生增加以及 CD133(+)细胞与人脐静脉内皮细胞(HUVEC)共培养时的分支点增加方面,与 BM MNC 相比,UCB CD133(+)细胞表现出更强的血管生成功能。

结论

总的来说,与 BM MNC 相比,UCB CD133(+)细胞在缺血反应中表现出更强的血管生成功能。

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