High-purity enrichment of functional cardiovascular cells from human iPS cells.
OBJECTIVE
A variety of human inherited heart diseases affect the normal functions of cardiomyocytes (CMs), endothelial cells (ECs), or smooth muscle cells (SMCs). To study human heart disease and generate cardiac cells for basic and translational research, an efficient strategy is needed for production of cardiac lineages from human stem cells. In the present study, a highly reproducible method was developed that can simultaneously enrich a large number of CMs and cardiac SMCs and ECs from human induced pluripotent stem (iPS) cells with high purity.


RESULTS
Human multipotent cardiovascular progenitor cells were generated from human iPS cells, followed by selective differentiation of the multipotent cardiovascular progenitor cells into CMs, ECs, and SMCs. With further fluorescence-activated cell sorting, each of the three cardiovascular cell types could be enriched with high purity (>90%). These enriched cardiovascular cells exhibited specific gene expression signatures and normal functions when assayed both in vitro and in vivo. Moreover, CMs purified from iPS cells derived from a patient with LEOPARD syndrome, a disease characterized by cardiac hypertrophy, showed the expected up-regulated expression of genes associated with cardiac hypertrophy.


CONCLUSIONS
Overall, our technical advance provides the means for generating a renewable resource of pure human cardiovascular cells that can be used to dissect the mechanisms of human inherited heart disease and for the future development of drug and cell therapeutics for heart disease.
Department of Developmental Biology, University of Pittsburgh School of Medicine, 530 45th Street, Rangos Research Center, Pittsburgh, PA 15201, USA.
Bo
B
Lin
Jong
J
Kim
Yanxin
Y
Li
Haiying
H
Pan
Xonia
X
Carvajal-Vergara
Guy
G
Salama
Tao
T
Cheng
Yong
Y
Li
Cecilia W
CW
Lo
Lei
L
Yang
D000818:Animals; D015703:Antigens, CD / Q000378:metabolism; D015415:Biomarkers / Q000378:metabolism; D006332:Cardiomegaly / Q000235:genetics / Q000473:pathology / Q000503:physiopathology; D015816:Cell Adhesion Molecules, Neuronal / Q000378:metabolism; D002454:Cell Differentiation; D019070:Cell Lineage; D002469:Cell Separation / Q000379:methods; D002478:Cells, Cultured; D005326:Fetal Proteins / Q000378:metabolism; D005434:Flow Cytometry; D005786:Gene Expression Regulation; D005838:Genotype; D006801:Humans; D057026:Induced Pluripotent Stem Cells / Q000378:metabolism / Q000502:physiology*; D044542:LEOPARD Syndrome / Q000235:genetics / Q000473:pathology / Q000503:physiopathology; D051379:Mice; D016688:Mice, Inbred NOD; D016513:Mice, SCID; D039902:Multipotent Stem Cells / Q000378:metabolism / Q000502:physiology*; D032383:Myocytes, Cardiac / Q000378:metabolism / Q000502:physiology*; D032389:Myocytes, Smooth Muscle / Q000378:metabolism / Q000502:physiology*; D010641:Phenotype; D012333:RNA, Messenger / Q000378:metabolism; D015203:Reproducibility of Results; D013997:Time Factors; D020410:Activated-Leukocyte Cell Adhesion Molecule
D015703:Antigens, CD; D015415:Biomarkers; D015816:Cell Adhesion Molecules, Neuronal; D005326:Fetal Proteins; D012333:RNA, Messenger; C105613:ALCAM protein, human; D020410:Activated-Leukocyte Cell Adhesion Molecule