Department of Chemical and Biological Engineering, University of Wisconsin, Madison, WI 53706, USA.
Proc Natl Acad Sci U S A. 2012 Jul 3;109(27):E1848-57. doi: 10.1073/pnas.1200250109. Epub 2012 May 29.
Human pluripotent stem cells (hPSCs) offer the potential to generate large numbers of functional cardiomyocytes from clonal and patient-specific cell sources. Here we show that temporal modulation of Wnt signaling is both essential and sufficient for efficient cardiac induction in hPSCs under defined, growth factor-free conditions. shRNA knockdown of β-catenin during the initial stage of hPSC differentiation fully blocked cardiomyocyte specification, whereas glycogen synthase kinase 3 inhibition at this point enhanced cardiomyocyte generation. Furthermore, sequential treatment of hPSCs with glycogen synthase kinase 3 inhibitors followed by inducible expression of β-catenin shRNA or chemical inhibitors of Wnt signaling produced a high yield of virtually (up to 98%) pure functional human cardiomyocytes from multiple hPSC lines. The robust ability to generate functional cardiomyocytes under defined, growth factor-free conditions solely by genetic or chemically mediated manipulation of a single developmental pathway should facilitate scalable production of cardiac cells suitable for research and regenerative applications.
人类多能干细胞 (hPSCs) 提供了从克隆和患者特异性细胞来源产生大量功能心肌细胞的潜力。在这里，我们表明在定义的、无生长因子的条件下，Wnt 信号的时间调节对于 hPSCs 中的有效心脏诱导是必不可少和充分的。在 hPSC 分化的初始阶段用 shRNA 敲低β-连环蛋白完全阻止了心肌细胞的特化，而此时糖原合成激酶 3 的抑制增强了心肌细胞的生成。此外，用糖原合成激酶 3 抑制剂顺序处理 hPSCs，然后诱导表达β-连环蛋白 shRNA 或 Wnt 信号化学抑制剂，可从多个 hPSC 系中产生高产量的几乎（高达 98%）纯功能的人类心肌细胞。仅通过遗传或化学介导的单一发育途径的操纵就能够在定义的、无生长因子的条件下产生功能心肌细胞的强大能力，应该有助于可扩展地生产适合研究和再生应用的心脏细胞。
