Selection of a common multipotent cardiovascular stem cell using the 3.4-kb MesP1 promoter fragment
Selection of a common multipotent cardiovascular stem cell using the 3.4-kb MesP1 promoter fragment
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DOI：
10.1007/s00395-012-0312-2
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发表时间：
2013-01-01
影响因子：
9.5
通讯作者：
Franz, Wolfgang-Michael
中科院分区：
文献类型：
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作者：
David, Robert;Schwarz, Florian;Franz, Wolfgang-Michael
常见的心血管祖细胞的特点和诱导的转录因子MesP 1的表达。为了表征这一群体，我们使用了我们小组先前描述的3.4 kb启动子片段。这用于通过基于截短的CD 4表面标记的磁性细胞分选从分化的ES干细胞中分离MesP 1阳性细胞。由于该近端启动子片段省略了远端非心血管生成增强子区域，我们能够获得高度富集的心血管祖细胞的同步部分。这导致约90%的细胞代表三种心血管谱系：心肌细胞、内皮细胞和平滑肌细胞，如蛋白质和mRNA分析所证实的。此外，心肌细胞组分的电生理学和药理学参数显示，几乎所有对应于分化第18天的多能早期/中期心肌细胞亚型。这些细胞的进一步分化没有受到损害，这从后期的强烈和同步跳动中可以看出。我们的工作有助于了解最早的心血管事件，并可能成为细胞治疗，组织工程和药理学测试中的培养皿使用多能干细胞衍生的，以及直接重编程的心血管细胞类型的重要先决条件。同样，这些细胞为大规模转录组和蛋白质组分析提供了理想的来源。
Common cardiovascular progenitor cells are characterized and induced by expression of the transcription factor MesP1. To characterize this population we used a 3.4-kb promoter fragment previously described by our group. This served to isolate MesP1-positive cells from differentiating ES stem cells via magnetic cell sorting based on a truncated CD4 surface marker. As this proximal promoter fragment omits a distal non-cardiovasculogenic enhancer region, we were able to achieve a synchronized fraction of highly enriched cardiovascular progenitors. These led to about 90 % of cells representing the three cardiovascular lineages: cardiomyocytes, endothelial cells and smooth muscle cells as evident from protein and mRNA analyses. In addition, electrophysiological and pharmacological parameters of the cardiomyocytic fraction show that almost all correspond to the multipotent early/intermediate cardiomyocyte subtype at day 18 of differentiation. Further differentiation of these cells was not impaired as evident from strong and synchronous beating at later stages. Our work contributes to the understanding of the earliest cardiovasculogenic events and may become an important prerequisite for cell therapy, tissue engineering and pharmacological testing in the culture dish using pluripotent stem cell-derived as well as directly reprogrammed cardiovascular cell types. Likewise, these cells provide an ideal source for large-scale transcriptome and proteome analyses.