Small molecule Wnt inhibitors enhance the efficiency of BMP-4-directed cardiac differentiation of human pluripotent stem cells.
Small molecule Wnt inhibitors enhance the efficiency of BMP-4-directed cardiac differentiation of human pluripotent stem cells.
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DOI：
10.1016/j.yjmcc.2011.04.012
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发表时间：
2011-09
影响因子：
5
通讯作者：
Qyang Y
中科院分区：
文献类型：
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作者：
Ren Y;Lee MY;Schliffke S;Paavola J;Amos PJ;Ge X;Ye M;Zhu S;Senyei G;Lum L;Ehrlich BE;Qyang Y
人类诱导多能干细胞（iPS）可能为产生患者特异性心肌细胞以研究心脏疾病机制和治疗提供独特的资源。然而，现有的从人iPS细胞生产心肌细胞的方法效率低下，限制了iPS细胞在基础和转化心脏研究中的应用。此外，需要准确记录iPS细胞衍生的心肌细胞动作电位时程（APD）变化的策略来监测APD相关的心脏疾病和快速药物筛选。我们研究了骨形态发生蛋白4（BMP-4）和Wnt/β-连环蛋白信号通路的调节是否可以诱导人iPS细胞的有效心脏分化。我们发现，与现有的分化策略相比，早期用BMP-4处理人iPS细胞，随后用小分子Wnt抑制剂进行后期处理，导致心肌细胞产量显著增加。使用免疫细胞化学染色和实时细胞内钙成像，我们发现这些诱导的心肌细胞表达典型的肌节标记物，表现出正常的节律性Ca 2+瞬变，并对β-肾上腺素能和电刺激作出反应。此外，人iPS细胞衍生的心肌细胞表现出响应于心脏活性药物普鲁卡因胺和维拉帕米的动作电位持续时间的特征性变化，使用电压敏感染料为基础的光学记录。因此，在人iPS细胞中BMP-4和Wnt信号通路的调节导致具有典型电生理功能和药理学响应性的心肌细胞的高效产生。使用人iPS细胞衍生的心肌细胞和应用钙和电压敏感染料直接，快速测量iPS细胞衍生的心肌细胞活性，有望为研究心脏疾病机制和治疗提供有吸引力的平台。
Human induced pluripotent stem (iPS) cells potentially provide a unique resource for generating patient-specific cardiomyocytes to study cardiac disease mechanisms and treatments. However, existing approaches to cardiomyocyte production from human iPS cells are inefficient, limiting the application of iPS cells in basic and translational cardiac research. Furthermore, strategies to accurately record changes in iPS cell-derived cardiomyocyte action potential duration (APD) are needed to monitor APD-related cardiac disease and for rapid drug screening. We examined whether modulation of the bone morphogenetic protein 4 (BMP-4) and Wnt/β-catenin signaling pathways could induce efficient cardiac differentiation of human iPS cells. We found that early treatment of human iPS cells with BMP-4 followed by late treatment with small molecule Wnt inhibitors led to a marked increase in production of cardiomyocytes compared to existing differentiation strategies. Using immunocytochemical staining and real-time intracellular calcium imaging, we showed that these induced cardiomyocytes expressed typical sarcomeric markers, exhibited normal rhythmic Ca2+ transients, and responded to both β-adrenergic and electric stimulation. Furthermore, human iPS cell-derived cardiomyocytes demonstrated characteristic changes in action potential duration in response to cardioactive drugs procainamide and verapamil using voltage-sensitive dye-based optical recording. Thus, modulation of the BMP-4 and Wnt signaling pathways in human iPS cells leads to highly efficient production of cardiomyocytes with typical electrophysiological function and pharmacologic responsiveness. The use of human iPS cell-derived cardiomyocytes and the application of calcium- and voltage-sensitive dyes for the direct, rapid measurement of iPS cell-derived cardiomyocyte activity promise to offer attractive platforms for studying cardiac disease mechanisms and therapeutics.
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影响因子：
64.8
作者：
Itzhaki, Ilanit;Maizels, Leonid;Gepstein, Lior
通讯作者：
Gepstein, Lior
影响因子：
64.8
作者：
Park, In-Hyun;Zhao, Rui;Daley, George Q.
通讯作者：
Daley, George Q.
影响因子：
20.1
作者：
Akar, FG;Rosenbaum, DS
通讯作者：
Rosenbaum, DS
DOI：
10.1073/pnas.0608156103
发表时间：
2006-11-14
影响因子：
11.1
作者：
Chen, Shuibing;Do, Jeong Tae;Ding, Sheng
通讯作者：
Ding, Sheng
影响因子：
64.8
作者：
通讯作者：
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