Bao Xiaoping, Lian Xiaojun, Hacker Timothy A, Schmuck Eric G, Qian Tongcheng, Bhute Vijesh J, Han Tianxiao, Shi Mengxuan, Drowley Lauren, Plowright Alleyn, Wang Qing-Dong, Goumans Marie-Jose, Palecek Sean P
Department of Chemical & Biological Engineering, University of Wisconsin, Madison, WI 53706, USA.
Departments of Biomedical Engineering, Biology and Huck Institutes of the Life Sciences, The Pennsylvania State University, University Park, PA 16802, USA.
Nat Biomed Eng. 2016;1. doi: 10.1038/s41551-016-0003. Epub 2016 Dec 5.
The epicardium contributes both multi-lineage descendants and paracrine factors to the heart during cardiogenesis and cardiac repair, underscoring its potential for cardiac regenerative medicine. Yet little is known about the cellular and molecular mechanisms that regulate human epicardial development and regeneration. Here, we show that the temporal modulation of canonical Wnt signaling is sufficient for epicardial induction from 6 different human pluripotent stem cell (hPSC) lines, including a WT1-2A-eGFP knock-in reporter line, under chemically-defined, xeno-free conditions. We also show that treatment with transforming growth factor beta (TGF-β)-signalling inhibitors permitted long-term expansion of the hPSC-derived epicardial cells, resulting in a more than 25 population doublings of WT1+ cells in homogenous monolayers. The hPSC-derived epicardial cells were similar to primary epicardial cells both and , as determined by morphological and functional assays, including RNA-seq. Our findings have implications for the understanding of self-renewal mechanisms of the epicardium and for epicardial regeneration using cellular or small-molecule therapies.
在心脏发生和心脏修复过程中，心外膜为心脏贡献了多谱系后代细胞和旁分泌因子，凸显了其在心脏再生医学中的潜力。然而，对于调节人类心外膜发育和再生的细胞及分子机制，我们却知之甚少。在此，我们表明，在化学成分明确、无动物源的条件下，经典Wnt信号通路的时序调控足以从6种不同的人类多能干细胞（hPSC）系中诱导出心外膜，其中包括一个WT1-2A-eGFP基因敲入报告细胞系。我们还表明，用转化生长因子β（TGF-β）信号抑制剂处理可使hPSC来源的心外膜细胞长期扩增，在均匀的单层细胞中，WT1+细胞的群体倍增超过25次。通过形态学和功能分析（包括RNA测序）确定，hPSC来源的心外膜细胞在[此处可能遗漏部分内容]方面与原代心外膜细胞相似。我们的研究结果对于理解心外膜的自我更新机制以及使用细胞或小分子疗法进行心外膜再生具有重要意义。
