Robust Generation of Cardiomyocytes from Human iPS Cells Requires Precise Modulation of BMP and WNT Signaling.
Robust Generation of Cardiomyocytes from Human iPS Cells Requires Precise Modulation of BMP and WNT Signaling.
复制标题
DOI：
10.1007/s12015-014-9564-6
复制
发表时间：
2015-08
影响因子：
4.8
通讯作者：
Edenhofer, Frank
中科院分区：
文献类型：
--
作者：
Kadari, Asifiqbal;Mekala, SubbaRao;Wagner, Nicole;Malan, Daniela;Koeth, Jessica;Doll, Katharina;Stappert, Laura;Eckert, Daniela;Peitz, Michael;Matthes, Jan;Sasse, Philipp;Herzig, Stefan;Bruestle, Oliver;Erguen, Suleyman;Edenhofer, Frank
关键词：
已经发表了各种能够使人类诱导多能干 (iPS) 细胞分化为心肌细胞的策略。然而，所涉及的信号通路的复杂性以及线间变异性损害了特定方案的应用，以从多个 iPS 系中稳健地获得心肌细胞。因此，有必要确定具有特定生长因子和小分子替代组合的优化方案，以增强心脏分化的稳健性。在这里，我们重点关注 BMP 和 WNT 信号传导的系统调节以增强心脏分化。此外，我们通过乳酸富集来提高心脏分化的功效。使用我们的方案，我们展示了从多个人类 iPS 系中有效衍生心肌细胞的方法。特别是，我们证明心肌细胞在 15 天内分化，通过心肌肌钙蛋白 T 流式细胞仪染色判断，分化效率高达 95%。通过 α-肌动蛋白染色、透射电子显微镜以及电生理分析对衍生的心肌细胞进行功能验证。我们希望我们的协议能够为功能性 iPS 细胞衍生的心肌细胞的规模化生产提供坚实的基础，这些心肌细胞可用于细胞替代疗法和疾病建模。本文的在线版本 (doi:10.1007/s12015-014-9564-6) 包含补充材料，可供授权用户使用。
Various strategies have been published enabling cardiomyocyte differentiation of human induced pluripotent stem (iPS) cells. However the complex nature of signaling pathways involved as well as line-to-line variability compromises the application of a particular protocol to robustly obtain cardiomyocytes from multiple iPS lines. Hence it is necessary to identify optimized protocols with alternative combinations of specific growth factors and small molecules to enhance the robustness of cardiac differentiation. Here we focus on systematic modulation of BMP and WNT signaling to enhance cardiac differentiation. Moreover, we improve the efficacy of cardiac differentiation by enrichment via lactate. Using our protocol we show efficient derivation of cardiomyocytes from multiple human iPS lines. In particular we demonstrate cardiomyocyte differentiation within 15 days with an efficiency of up to 95 % as judged by flow cytometry staining against cardiac troponin T. Cardiomyocytes derived were functionally validated by alpha-actinin staining, transmission electron microscopy as well as electrophysiological analysis. We expect our protocol to provide a robust basis for scale-up production of functional iPS cell-derived cardiomyocytes that can be used for cell replacement therapy and disease modeling. The online version of this article (doi:10.1007/s12015-014-9564-6) contains supplementary material, which is available to authorized users.
登录
查看更多内容
影响因子：
3.7
作者：
Fuerer C;Nusse R
通讯作者：
Nusse R
影响因子：
23.9
作者：
Burridge, Paul W.;Keller, Gordon;Gold, Joseph D.;Wu, Joseph C.
通讯作者：
Wu, Joseph C.
影响因子：
168.9
作者：
Lopez, Alan D.;Mathers, Colin D.;Murray, Christopher J. L.
通讯作者：
Murray, Christopher J. L.
影响因子：
64.8
作者：
Laugwitz, KL;Moretti, A;Chien, KR
通讯作者：
Chien, KR
影响因子：
64.8
作者：
Laflamme, Michael A.;Murry, Charles E.
通讯作者：
Murry, Charles E.