Highly efficient induction and long-term maintenance of multipotent cardiovascular progenitors from human pluripotent stem cells under defined conditions
Highly efficient induction and long-term maintenance of multipotent cardiovascular progenitors from human pluripotent stem cells under defined conditions
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在特定条件下高效诱导和长期维持人类多能干细胞的多能心血管祖细胞
DOI：
10.1038/cr.2013.102
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发表时间：
2013-09-01
期刊：
影响因子：
44.1
通讯作者：
Yang, Huang-Tian
中科院分区：
文献类型：
--
作者：
Cao, Nan;Liang, He;Yang, Huang-Tian
由人胚胎干细胞（hESCs）和人诱导多能干细胞（hiPSCs）衍生而来的心血管祖细胞（CVPCs）在心血管发育和心脏疾病的细胞治疗研究中具有广阔的应用前景，但其高效生成和稳定维持的困难给其应用带来了挑战。本研究旨在通过调节涉及人类心血管规范和CVPC自我更新的关键早期发育途径，开发化学定义的系统，用于hpsc衍生的CVPC的稳健生成和稳定繁殖。在此，我们报告了骨形态发生蛋白4 （BMP4）、糖原合成酶激酶3 （GSK3）抑制剂CHIR99021和抗坏血酸的组合足以在化学定义的培养基中，在无饲料和无血清的培养条件下，将单层培养的hPSCs（包括hESCs和hiPSCs）快速转化为均匀的CVPCs。在无饲料和无血清条件下，这些CVPCs稳定地自我更新，当BMP、GSK3和Activin/Nodal途径的诱导分化信号同时消除时，这些CVPCs扩增超过10.7倍。此外，这些CVPCs表现出预期的全基因组CVPCs分子特征，保留了在体外产生主要心血管细胞谱系的潜力，包括心肌细胞、平滑肌细胞和内皮细胞，并且在体内无致瘤性。总之，本文报道的已建立的系统允许高效生成和稳定维持hpsc衍生的CVPCs，这是研究早期胚胎心血管发育的有力工具，并为心肌再生医学提供了潜在的安全细胞来源。
Cardiovascular progenitor cells (CVPCs) derived from human pluripotent stem cells (hPSCs), including human embryonic stem cells (hESCs) and human induced pluripotent stem cells (hiPSCs), hold great promise for the study of cardiovascular development and cell-based therapy of heart diseases, but their applications are challenged by the difficulties in their efficient generation and stable maintenance. This study aims to develop chemically defined systems for robust generation and stable propagation of hPSC-derived CVPCs by modulating the key early developmental pathways involved in human cardiovascular specification and CVPC self-renewal. Herein we report that a combination of bone morphogenetic protein 4 (BMP4), glycogen synthase kinase 3 (GSK3) inhibitor CHIR99021 and ascorbic acid is sufficient to rapidly convert monolayer-cultured hPSCs, including hESCs and hiPSCs, into homogeneous CVPCs in a chemically defined medium under feeder-and serum-free culture conditions. These CVPCs stably self-renewed under feeder-and serum-free conditions and expanded over 10 7-fold when the differentiation-inducing signals from BMP, GSK3 and Activin/Nodal pathways were simultaneously eliminated. Furthermore, these CVPCs exhibited expected genome-wide molecular features of CVPCs, retained potentials to generate major cardiovascular lineages including cardiomyocytes, smooth muscle cells and endothelial cells in vitro, and were non-tumorigenic in vivo. Altogether, the established systems reported here permit efficient generation and stable maintenance of hPSC-derived CVPCs, which represent a powerful tool to study early embryonic cardiovascular development and provide a potentially safe source of cells for myocardial regenerative medicine.