An engineered cardiac reporter cell line identifies human embryonic stem cell-derived myocardial precursors

• Published in: PloS one 2011-01, Vol.6 (1), p.e16004
• Main Authors: Ritner, Carissa, Wong, Sharon S Y, King, Frank W, Mihardja, Shirley S, Liszewski, Walter, Erle, David J, Lee, Randall J, Bernstein, Harold S
• Format: Article
• Language: English
• Subjects: Analysis; Antigens; Bayesian analysis; Binding sites; Bioinformatics; Biology; Bone morphogenetic proteins; Cardiomyocytes; Cell Line; Conduction; Coronary artery disease; Drug resistance; Embryo cells; Embryonic stem cells; Embryonic Stem Cells - cytology; Embryos; Gene expression; Gene Expression Profiling; Genomes; Green Fluorescent Proteins; Heart; Heart diseases; Humans; Immunocytochemistry; Journalists; Localization; Medical research; Medical treatment; Medicine; Microelectrodes; Muscle proteins; Myoblasts, Cardiac - cytology; Myosin; Myosin Heavy Chains; Organs; Pathways; Precursors; Protein Engineering - methods; Signal Transduction - genetics; Signaling; Smooth muscle; Stem cells; Teratoma; Tissues; Transforming growth factor; Transforming Growth Factor beta; Transforming growth factor-b; Ventricle; Ventricular Myosins; Wnt protein; Wnt Proteins
• ISSN: 1932-6203; 1932-6203
• DOI: 10.1371/journal.pone.0016004

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.