A novel in vitro model system for smooth muscle differentiation from human embryonic stem cell-derived mesenchymal cells
A novel in vitro model system for smooth muscle differentiation from human embryonic stem cell-derived mesenchymal cells
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DOI：
10.1152/ajpcell.00298.2012
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发表时间：
2013-02-01
影响因子：
5.5
通讯作者：
Chen, Shi-You
中科院分区：
文献类型：
--
作者：
Guo, Xia;Stice, Steven L.;Chen, Shi-You
郭X，Stice SL，Boyd NL，Chen SY。人胚胎干细胞来源的间充质细胞向平滑肌分化的一种新的体外模型系统。Am J Physiol Cell Physiol 304：C289-C298，2013.首次发表于2012年12月5日; doi：10.1152/ajpcell.00298.2012.-本研究的目的是建立一种新的人胚胎干细胞来源的间充质细胞（hES-MCs）向平滑肌细胞（SMC）分化的体外模型。我们发现，hES-MCs分化为SMC的转化生长因子-β（TGF-β）的剂量和时间依赖性的方式，表现为SMC特异性基因平滑肌α-肌动蛋白，钙调蛋白，平滑肌肌球蛋白重链的表达。然而，在正常生长条件下，hES-MCs的分化能力非常有限。hES-MC衍生的SMC具有细长的纺锤形形态，并响应于卡巴胆碱和KCl的诱导而收缩。KCl诱导的钙瞬变在这些细胞中也很明显。与亲本细胞相比，TGF-β处理的hES-MCs由于持续的SMC表型而维持了更长时间的内皮管形成。TGF-β诱导分化的机制是Smad和血清反应因子/心肌素依赖性的。TGF-β通过多种信号途径调节心肌蛋白的表达，包括Smad 2/3、p38 MAPK和PI 3 K。重要的是，我们发现在SMC谱系确定之前，中胚层中存在低水平的myocardin，并且直到分化过程开始才诱导高水平的myocardin。总之，我们的研究特征在于一种新的SMC分化模型，可用于研究血管发育过程中胚层的人SMC分化。
Guo X, Stice SL, Boyd NL, Chen SY. A novel in vitro model system for smooth muscle differentiation from human embryonic stem cell-derived mesenchymal cells. Am J Physiol Cell Physiol 304: C289-C298, 2013. First published December 5, 2012; doi:10.1152/ajpcell.00298.2012.-The objective of this study was to develop a novel in vitro model for smooth muscle cell (SMC) differentiation from human embryonic stem cell-derived mesenchymal cells (hES-MCs). We found that hES-MCs were differentiated to SMCs by transforming growth factor-beta (TGF-beta) in a dose-and time-dependent manner as demonstrated by the expression of SMC-specific genes smooth muscle alpha-actin, calponin, and smooth muscle myosin heavy chain. Under normal growth conditions, however, the differentiation capacity of hES-MCs was very limited. hES-MC-derived SMCs had an elongated and spindle-shaped morphology and contracted in response to the induction of carbachol and KCl. KCl-induced calcium transient was also evident in these cells. Compared with the parental cells, TGF-beta-treated hES-MCs sustained the endothelial tube formation for a longer time due to the sustained SMC phenotype. Mechanistically, TGF-beta-induced differentiation was both Smad-and serum response factor/myocardin dependent. TGF-beta regulated myocardin expression via multiple signaling pathways including Smad2/3, p38 MAPK, and PI3K. Importantly, we found that a low level of myocardin was present in mesoderm prior to SMC lineage determination, and a high level of myocardin was not induced until the differentiation process was initiated. Taken together, our study characterized a novel SMC differentiation model that can be used for studying human SMC differentiation from mesoderm during vascular development.