Highly efficient induction and long-term maintenance of multipotent cardiovascular progenitors from human pluripotent stem cells under defined conditions

• Published in: Cell research Vol. 23; no. 9; pp. 1119 - 1132
• Main Authors: Cao, Nan, Liang, He, Huang, Jijun, Wang, Jia, Chen, Yixiong, Chen, Zhongyan, Yang, Huang-Tian
• Format: Journal Article
• Language: English
• Published: London Nature Publishing Group UK 01.09.2013; Nature Publishing Group
• Subjects: 631/136/532/1360; 631/136/532/2064; 631/1647/1407/651; 692/699/75; Ascorbic Acid - pharmacology; Biomedical and Life Sciences; Bone Morphogenetic Protein 4 - pharmacology; Cardiovascular diseases; Cell Biology; Cell Culture Techniques; Cell Differentiation - drug effects; Cell- and Tissue-Based Therapy; Cells, Cultured; Embryonic Stem Cells - drug effects; Embryonic Stem Cells - metabolism; Glycogen Synthase Kinase 3 - antagonists & inhibitors; Heart - embryology; Heart Diseases - therapy; Humans; Induced Pluripotent Stem Cells - drug effects; Induced Pluripotent Stem Cells - metabolism; Life Sciences; Myocardium - cytology; Original; original-article; Pyridines - pharmacology; Pyrimidines - pharmacology; Stem cells; 人类胚胎干细胞; 分化诱导; 多能干细胞; 心血管; 祖细胞; 自我更新; 血管发育; 骨形态发生蛋白; directed differentiation; human pluripotent stem cells; cardiovascular progenitor cells; chemically defined medium; progenitor maintenance
• ISSN: 1001-0602; 1748-7838; 1748-7838
• DOI: 10.1038/cr.2013.102

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 devel- opmental 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 homoge- neous 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 107-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.