Induction and enhancement of cardiac cell differentiation from mouse and human induced pluripotent stem cells with cyclosporin-A.
Induction and enhancement of cardiac cell differentiation from mouse and human induced pluripotent stem cells with cyclosporin-A.
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DOI：
10.1371/journal.pone.0016734
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发表时间：
2011-02-22
期刊：
影响因子：
3.7
通讯作者：
Yamashita JK
中科院分区：
文献类型：
--
作者：
Fujiwara M;Yan P;Otsuji TG;Narazaki G;Uosaki H;Fukushima H;Kuwahara K;Harada M;Matsuda H;Matsuoka S;Okita K;Takahashi K;Nakagawa M;Ikeda T;Sakata R;Mummery CL;Nakatsuji N;Yamanaka S;Nakao K;Yamashita JK
诱导性多能干细胞（Induced pluripotent stem cells，iPSCs）是一种通过重编程从成年小鼠和人体组织中分离的新型干细胞。阐明其分化为功能性心肌细胞的机制和探索有效的方法对于开发心脏细胞模型和未来的再生疗法至关重要。我们之前建立了一种新的小鼠胚胎干细胞（ESC）和iPSC分化系统，其中心血管细胞可以从Flk 1+共同祖细胞系统诱导，并鉴定出高度心源性祖细胞为Flk 1 +/CXCR 4 +/VE-钙粘蛋白-（FCV）细胞。我们还报道了环孢菌素A（CSA）显著增加小鼠ESC的FCV祖细胞和心肌细胞诱导。在这里，我们结合了这些技术，并将其扩展到小鼠和人类iPSC。纯化的小鼠iPSC衍生的Flk 1+细胞与0 P9基质细胞的共培养诱导心肌细胞分化，而向Flk 1+细胞添加CSA显著增加心肌细胞和FCV祖细胞分化。通过与END-2内脏内胚层样细胞共培养从人iPSC获得自发搏动集落。通过在中胚层阶段添加CSA，来自人iPSC的搏动集落的外观增加约4.3倍。CSA扩增的人iPSC衍生的心肌细胞显示出心肌细胞的各种心脏标志物表达、同步钙瞬变、心肌细胞样动作电位、药理学反应和超微结构特征。这些结果为从iPSCs中获得功能性心肌细胞提供了技术基础。
Induced pluripotent stem cells (iPSCs) are novel stem cells derived from adult mouse and human tissues by reprogramming. Elucidation of mechanisms and exploration of efficient methods for their differentiation to functional cardiomyocytes are essential for developing cardiac cell models and future regenerative therapies. We previously established a novel mouse embryonic stem cell (ESC) and iPSC differentiation system in which cardiovascular cells can be systematically induced from Flk1+ common progenitor cells, and identified highly cardiogenic progenitors as Flk1+/CXCR4+/VE-cadherin− (FCV) cells. We have also reported that cyclosporin-A (CSA) drastically increases FCV progenitor and cardiomyocyte induction from mouse ESCs. Here, we combined these technologies and extended them to mouse and human iPSCs. Co-culture of purified mouse iPSC-derived Flk1+ cells with OP9 stroma cells induced cardiomyocyte differentiation whilst addition of CSA to Flk1+ cells dramatically increased both cardiomyocyte and FCV progenitor cell differentiation. Spontaneously beating colonies were obtained from human iPSCs by co-culture with END-2 visceral endoderm-like cells. Appearance of beating colonies from human iPSCs was increased approximately 4.3 times by addition of CSA at mesoderm stage. CSA-expanded human iPSC-derived cardiomyocytes showed various cardiac marker expressions, synchronized calcium transients, cardiomyocyte-like action potentials, pharmacological reactions, and ultra-structural features as cardiomyocytes. These results provide a technological basis to obtain functional cardiomyocytes from iPSCs.
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影响因子：
20.1
作者：
Reinecke H;Minami E;Zhu WZ;Laflamme MA
通讯作者：
Laflamme MA
影响因子：
4.8
作者：
Moretti, Alessandra;Bellin, Milena;Laugwitz, Karl-Ludwig
通讯作者：
Laugwitz, Karl-Ludwig
影响因子：
5.2
作者：
Passier, R;Oostwaard, DWV;Mummery, C
通讯作者：
Mummery, C
影响因子：
64.5
作者：
Takahashi, Kazutoshi;Yamanaka, Shinya
通讯作者：
Yamanaka, Shinya
影响因子：
15.9
作者：
Kehat, I;Kenyagin-Karsenti, D;Gepstein, L
通讯作者：
Gepstein, L