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.