Differentiation, survival, and function of embryonic stem cell-derived endothelial cells for ischemic heart disease
Differentiation, survival, and function of embryonic stem cell-derived endothelial cells for ischemic heart disease
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DOI：
10.1161/circulationaha.106.680561
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发表时间：
2007-09-11
期刊：
影响因子：
37.8
通讯作者：
Wu, Joseph C.
中科院分区：
文献类型：
--
作者：
Li, Zongjin;Wu, Jenny C.;Wu, Joseph C.
背景-胚胎干细胞（ES）以其自我更新和多能性的能力而闻名。在这里，我们的特点分化的ES细胞衍生的内皮细胞（ESC-ECs），使用分子成像技术来检查他们的生存在体内，并确定治疗效果的ESC-ECs为缺血性injuries.Methods和Results恢复心脏功能-小鼠ES细胞转染的血管内皮钙粘蛋白启动子驱动增强的绿色荧光蛋白（pVE-钙粘蛋白-eGFP）组成的结构。使用Flk-1（第4天的早期EC标志物）和VE-钙粘蛋白（第8天的晚期EC标志物）通过FACS分析检测ES细胞向EC的分化。分离后，这些ESC-EC表达与成年小鼠肺内皮细胞相似的内皮细胞标记物，形成血管样通道，并掺入DiI标记的乙酰化低密度脂蛋白（DiI-Ac-LDL）。对于体内成像，用驱动萤火虫荧光素酶和单体红色荧光蛋白（pUb-Fluc-mRFP）的泛素启动子转导ES细胞。通过离体成像分析（R-2 = 0.98）和通过体外酶测定（R-2 = 0.94），Fluc信号和细胞数量之间存在稳健的相关性。随后，将5 x 10（5）个ESC-EC或PBS（作为对照）注射到接受LAD结扎的小鼠心脏中（每组n = 15）。生物发光成像显示移植的ESC-ECs的纵向存活约8周。超声心动图显示，与对照组相比，ESC-EC组的功能显著改善（P = 0.04）。最后，尸检分析证实增加存在的小毛细血管和小静脉在梗死zones by CD 31 staining.Conclusions -这是第一个研究跟踪移植的ESC-ECs在心脏的命运和功能。随着进一步的验证，这些ESC-EC可能成为一个有价值的来源，诱导血管生成的细胞治疗心肌缺血。
Background - Embryonic stem (ES) cells are distinguished by their capacity for self-renewal and pluripotency. Here we characterize the differentiation of ES cell-derived endothelial cells (ESC-ECs), use molecular imaging techniques to examine their survival in vivo, and determine the therapeutic efficacy of ESC-ECs for restoration of cardiac function after ischemic injury.Methods and Results - Murine ES cells were transfected with a construct composed of a vascular endothelial cadherin promoter driving enhanced green fluorescence protein (pVE-cadherin-eGFP). Differentiation of ES cells to ECs was detected by FACS analysis using Flk-1 (early EC marker at day 4) and VE-cadherin (late EC marker at day 8). After isolation, these ESC-ECs express endothelial cell markers similar to adult mouse lung endothelial cells, form vascular-like channels, and incorporate DiI-labeled acetylated low-density lipoprotein (DiI-Ac-LDL). For in vivo imaging, ES cells were transduced with an ubiquitin promoter driving firefly luciferase and monomeric red fluorescence protein (pUb-Fluc-mRFP). A robust correlation exists between Fluc signals and cell numbers by ex vivo imaging analysis (R-2 = 0.98) and by in vitro enzyme assay (R-2 = 0.94). Afterward, 5 x 10(5) ESC-ECs or PBS (as control) was injected into the hearts of mice undergoing LAD ligation (n = 15 per group). Bioluminescence imaging showed longitudinal survival of transplanted ESC-ECs for approximate to 8 weeks. Echocardiogram demonstrated significant functional improvement in the ESC-EC group compared with control (P = 0.04). Finally, postmortem analysis confirmed increased presence of small capillaries and venules in the infarcted zones by CD31 staining.Conclusions - This is the first study to track the fate and function of transplanted ESC-ECs in the heart. With further validation, these ESC-ECs could become a valuable source of cell therapy for induction of angiogenesis in the treatment of myocardial ischemia.