Prospective identification of cardiac progenitors by a novel single cell-based cardiomyocyte induction
Prospective identification of cardiac progenitors by a novel single cell-based cardiomyocyte induction
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DOI：
10.1096/fj.04-3540fje
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发表时间：
2005-07-01
期刊：
影响因子：
4.8
通讯作者：
Nishikawa, SI
中科院分区：
文献类型：
--
作者：
Yamashita, JK;Takano, M;Nishikawa, SI
在细胞水平上对心肌细胞分化过程的研究是心脏发育和再生研究中不可缺少的。以前，我们已经建立了一个胚胎干细胞分化系统，再现早期血管发育的祖细胞表达Flk 1，血管内皮生长因子受体，通过二维培养和流式细胞术的组合应用。在这里，我们表明，心肌细胞可以成功地诱导从一个单一的Flk 1（+）细胞的二维培养，使分化的心肌细胞的直接观察和心脏祖细胞的潜力的前瞻性识别。Flk 1+细胞与OP 9基质细胞共培养后，可从单一细胞分化为心肌细胞和内皮细胞，且不依赖于融合。从Flk 1+细胞到心肌细胞的中间阶段的细胞群中，Flk 1（+）/CXCR 4（+）/血管内皮钙粘素（-）细胞在单细胞水平上是心脏特异性祖细胞。骨形态发生蛋白抑制剂Noggin通过抑制心肌祖细胞的诱导而阻断心肌细胞的分化。然而，wnt抑制剂Dkk-1或Frizzled-8/Fc嵌合蛋白增强，但wnt 3a抑制，心肌细胞分化。心肌细胞分化过程的体外复制将为从细胞和分子水平阐明心脏发育过程提供有力的工具，为心脏再生提供多种靶点。
Dissection of cardiomyocyte differentiation process at the cellular level is indispensable in the research for cardiac development and regeneration. Previously, we have established an embryonic stem cell differentiation system that reproduces early vascular development from progenitor cells that express Flk1, a vascular endothelial growth factor receptor, by the combinatory application of 2-dimensional culture and flowcytometry. Here we show that cardiomyocytes can be successfully induced from a single Flk1(+) cell on 2-dimensional culture, enabling the direct observation of differentiating cardiomyocytes and the prospective identification of cardiac progenitor potentials. Flk1+ cells could give rise to cardiomyocytes, as well as endothelial cells, from a single cell by the co-culture on OP9 stroma cells in a fusion-independent manner. Among the cell populations in intermediate stages from Flk1+ cells to cardiomyocytes, Flk1(+)/CXCR4(+)/ vascular endothelial cadherin(-) cells were cardiac-specific progenitors at the single cell level. Noggin, a bone morphogenetic protein inhibitor, abolished cardiomyocyte differentiation by inhibiting the cardiac progenitor induction. However, wnt inhibitors Dkk-1 or Frizzled-8/Fc chimeric protein augmented, but wnt3a inhibited, cardiomyocyte differentiation. In vitro reproduction of cardiomyocyte differentiation process should be a potent tool for the cellular and molecular elucidation of cardiac development, which would provide various targets for cardiac regeneration.