Human cardiovascular progenitor cells develop from a KDR plus embryonic-stem-cell-derived population
Human cardiovascular progenitor cells develop from a KDR plus embryonic-stem-cell-derived population
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DOI：
10.1038/nature06894
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发表时间：
2008-05-22
期刊：
影响因子：
64.8
通讯作者：
Keller, Gordon M.
中科院分区：
文献类型：
--
作者：
Yang, Lei;Soonpaa, Mark H.;Keller, Gordon M.
功能心脏由不同的中胚层细胞组成，包括心肌细胞、内皮细胞和血管平滑肌细胞。对小鼠胚胎和小鼠胚胎干细胞分化模型的研究提供了证据，表明这三个谱系是从一个共同的Flk-1(+)（激酶插入结构域蛋白受体，也称为Kdr）心血管祖细胞（代表中胚层规范的最早阶段之一）发展到心血管谱系的(1)。为了确定在人类心脏发生过程中是否存在类似的祖细胞，我们分析了人类胚胎干细胞分化培养中心血管谱系的发展。本研究表明，在无血清培养基中，激活素A、骨形态发生蛋白4 （BMP4）、碱性成纤维细胞生长因子（bFGF，也称为FGF2）、血管内皮生长因子（VEGF，也称为VEGFA）和dickkopf同源物1 （DKK1）联合诱导后，人胚胎干细胞衍生的胚胎样体产生KDRlow/C-KIT (CD117)（阴性）群体，在体外和移植后，在体内显示心脏、内皮和血管平滑肌电位。在单层培养中，这些KDR低/C-KIT阴性细胞分化产生由大于50%的收缩心肌细胞组成的群体。来自KDRlow/C-KITneg部分的群体在甲基纤维素培养物中培养时产生包含所有三种谱系的菌落。限制性稀释研究和细胞混合实验的结果支持这些集落是克隆的解释，表明它们是从心血管集落形成细胞发育而来的。总之，这些发现确定了人类心血管祖先，确定了人类心脏发育的最早阶段之一。
The functional heart is comprised of distinct mesoderm-derived lineages including cardiomyocytes, endothelial cells and vascular smooth muscle cells. Studies in the mouse embryo and the mouse embryonic stem cell differentiation model have provided evidence indicating that these three lineages develop from a common Flk-1(+) (kinase insert domain protein receptor, also known as Kdr) cardiovascular progenitor that represents one of the earliest stages in mesoderm specification to the cardiovascular lineages(1). To determine whether a comparable progenitor is present during human cardiogenesis, we analysed the development of the cardiovascular lineages in human embryonic stem cell differentiation cultures. Here we show that after induction with combinations of activin A, bone morphogenetic protein 4 (BMP4), basic fibroblast growth factor (bFGF, also known as FGF2), vascular endothelial growth factor (VEGF, also known as VEGFA) and dickkopf homolog 1 (DKK1) in serum-free media, human embryonic-stem-cell-derived embryoid bodies generate a KDRlow/C-KIT (CD117)(neg) population that displays cardiac, endothelial and vascular smooth muscle potential in vitro and, after transplantation, in vivo. When plated in monolayer cultures, these KDR low/C-KIT neg cells differentiate to generate populations consisting of greater than 50% contracting cardiomyocytes. Populations derived from the KDRlow/C-KITneg fraction give rise to colonies that contain all three lineages when plated in methylcellulose cultures. Results from limiting dilution studies and cell-mixing experiments support the interpretation that these colonies are clones, indicating that they develop from a cardiovascular colonyforming cell. Together, these findings identify a human cardiovascular progenitor that defines one of the earliest stages of human cardiac development.