A Human iPSC Double-Reporter System Enables Purification of Cardiac Lineage Subpopulations with Distinct Function and Drug Response Profiles
A Human iPSC Double-Reporter System Enables Purification of Cardiac Lineage Subpopulations with Distinct Function and Drug Response Profiles
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DOI：
10.1016/j.stem.2019.02.015
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发表时间：
2019-05-02
期刊：
影响因子：
23.9
通讯作者：
Wu, Joseph C.
中科院分区：
文献类型：
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作者：
Zhang, Joe Z.;Termglinchan, Vittavat;Wu, Joseph C.
心脏谱系的多样性导致了人诱导多能干细胞(HiPSC)来源的心肌细胞(CMS)的异质性。在这里，我们报道了HiPSC TBX5 Clover2和NKX2-5 TagRFP双报告基因的产生，以描绘心脏谱系并分离谱系特异性亚群。分子分析表明，根据TBX5和NKX2-5、TBX5+NKX2-5+、TBX5+NKX2-5-、TBX5-NKX2-5+和TBX5-NKX2-5-的差异表达，可以分离出四个不同的亚群，分别模拟第一心域、心外膜、第二心域和内皮谱系。基因和功能特征表明，每个亚群都分化为特定的心肌细胞。我们进一步确定Corin作为分离TBX5+NKX2-5+亚群的细胞表面标记，并展示了谱系特异性CMS用于精确的药物测试。我们预计，该工具将有助于研究心脏谱系的规范和分离特定的心脏亚群，用于药物筛选、组织工程和疾病建模。
The diversity of cardiac lineages contributes to the heterogeneity of human induced pluripotent stem cell (hiPSC)-derived cardiomyocytes (CMs). Here, we report the generation of a hiPSC TBX5 Clover2 and NKX2-5 TagRFP double reporter to delineate cardiac lineages and isolate lineage-specific subpopulations. Molecular analyses reveal that four different subpopulations can be isolated based on the differential expression of TBX5 and NKX2-5, TBX5+NKX2-5+, TBX5+NKX2-5-, TBX5-NKX2-5+, and TBX5-NKX2-5-,mimicking the first heart field, epicardial, second heart field, and endothelial lineages, respectively. Genetic and functional characterization indicates that each subpopulation differentiates into specific cardiac cells. We further identify CORIN as a cell-surface marker for isolating the TBX5+NKX2-5+ subpopulation and demonstrate the use of lineage-specific CMs for precise drug testing. We anticipate that this tool will facilitate the investigation of cardiac lineage specification and isolation of specific cardiac subpopulations for drug screening, tissue engineering, and disease modeling.