An engineered cardiac reporter cell line identifies human embryonic stem cell-derived myocardial precursors.
Unlike some organs, the heart is unable to repair itself after injury. Human embryonic stem cells (hESCs) grow and divide indefinitely while maintaining the potential to develop into many tissues of the body. As such, they provide an unprecedented opportunity to treat human diseases characterized by tissue loss. We have identified early myocardial precursors derived from hESCs (hMPs) using an α-myosin heavy chain (αMHC)-GFP reporter line. We have demonstrated by immunocytochemistry and quantitative real-time PCR (qPCR) that reporter activation is restricted to hESC-derived cardiomyocytes (CMs) differentiated in vitro, and that hMPs give rise exclusively to muscle in an in vivo teratoma formation assay. We also demonstrate that the reporter does not interfere with hESC genomic stability. Importantly, we show that hMPs give rise to atrial, ventricular and specialized conduction CM subtypes by qPCR and microelectrode array analysis. Expression profiling of hMPs over the course of differentiation implicate Wnt and transforming growth factor-β signaling pathways in CM development. The identification of hMPs using this αMHC-GFP reporter line will provide important insight into the pathways regulating human myocardial development, and may provide a novel therapeutic reagent for the treatment of cardiac disease.
Cardiovascular Research Institute, University of California San Francisco, San Francisco, California, United States of America.
Carissa
C
Ritner
Sharon S Y
SS
Wong
Frank W
FW
King
Shirley S
SS
Mihardja
Walter
W
Liszewski
David J
DJ
Erle
Randall J
RJ
Lee
Harold S
HS
Bernstein
D002460:Cell Line; D053595:Embryonic Stem Cells / Q000166:cytology*; D020869:Gene Expression Profiling; D049452:Green Fluorescent Proteins; D006801:Humans; D032386:Myoblasts, Cardiac / Q000166:cytology*; D018995:Myosin Heavy Chains; D015202:Protein Engineering / Q000379:methods*; D015398:Signal Transduction / Q000235:genetics; D016212:Transforming Growth Factor beta; D024723:Ventricular Myosins; D051153:Wnt Proteins
D016212:Transforming Growth Factor beta; D051153:Wnt Proteins; D049452:Green Fluorescent Proteins; D024723:Ventricular Myosins; D018995:Myosin Heavy Chains
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