Key Laboratory of Stem Cell Biology, Institute of Health Sciences, Shanghai Institutes for Biological Sciences, Chinese Academy of Sciences & Shanghai Jiao Tong University School of Medicine (SJTUSM), Shanghai 200025, China.
Cell Res. 2013 Sep;23(9):1119-32. doi: 10.1038/cr.2013.102. Epub 2013 Jul 30.
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.
人心血管祖细胞（CVPCs）来源于人类多能干细胞（hPSCs），包括人类胚胎干细胞（hESCs）和人类诱导多能干细胞（hiPSCs），在心血管发育和基于细胞的心脏病治疗方面具有广阔的应用前景，但它们的应用受到高效生成和稳定维持的困难的挑战。本研究旨在通过调节涉及人心血管特化和 CVPC 自我更新的关键早期发育途径，开发化学定义系统来稳健地生成和稳定扩增 hPSC 衍生的 CVPCs。在此，我们报告称，骨形态发生蛋白 4（BMP4）、糖原合酶激酶 3（GSK3）抑制剂 CHIR99021 和抗坏血酸的组合足以在无饲养细胞和血清的培养条件下，在化学定义的培养基中从单层培养的 hPSCs（包括 hESCs 和 hiPSCs）快速转化为同质的 CVPCs。这些 CVPCs 在无饲养细胞和血清的条件下稳定自我更新，当 BMP、GSK3 和激活素/诺达途径的分化诱导信号同时消除时，可扩增超过 107 倍。此外，这些 CVPCs表现出预期的 CVPC 全基因组分子特征，在体外保留生成主要心血管谱系（包括心肌细胞、平滑肌细胞和内皮细胞）的潜力，并且在体内无致瘤性。总之，本文建立的系统可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安全的细胞来源。
