Coexpression of platelet-derived growth factor receptor alpha and fetal liver kinase 1 enhances cardiogenic potential in embryonic stem cell differentiation in vitro

SUMMARYThe GATA4 transcription factor is implicated in promoting cardiogenesis in combination with other factors, including TBX5, MEF2C and BAF60C. However, when expressed in embryonic stem cells (ESCs), GATA4 was shown to promote endoderm, not cardiac mesoderm. The capacity of related GATA factors to promote cardiogenesis is untested. We found that expression of the highly related gene, Gata5, very efficiently promotes cardiomyocyte fate from murine ESCs. Gata5 directs development of beating sheets of cells that express cardiac troponin T and show a full range of action potential morphologies that are responsive to pharmacological stimulation. We discovered that by removing serum from the culture conditions, GATA4 and GATA6 are each also able to efficiently promote cardiogenesis in ESC derivatives, with some distinctions. Thus, GATA factors can function in ESC derivatives upstream of other cardiac transcription factors to direct the efficient generation of cardiomyocytes.

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“…Both the cell beat rate (Fig. 2F) ( Hirata et al, 2007;Kattman et al, 2011 (Kattman et al, 2011). Each population was isolated by FACS (Fig.…”

Section: Research Reportmentioning

confidence: 99%

GATA factors efficiently direct cardiac fate from embryonic stem cells

et al. 2013

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“…A number of ESC-CPs have been identified and characterized (10,26,27). These cells have been isolated at different efficiencies using transcription factor-driven promoters, or using cell surface expressed antigens.…”

Section: Injection Of Esc-derived Cps Improves the Electrical Functiomentioning

confidence: 99%

Interrogating functional integration between injected pluripotent stem cell-derived cells and surrogate cardiac tissue

Song¹,

Yoon²,

Kattman

et al. 2009

Proc. Natl. Acad. Sci. U.S.A.

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Myocardial infarction resulting in irreversible loss of cardiomyocytes (CMs) remains a leading cause of heart failure. Although cell transplantation has modestly improved cardiac function, major challenges including increasing cell survival, engraftment, and functional integration with host tissue, remain. Embryonic stem cells (ESCs), which can be differentiated into cardiac progenitors (CPs) and CMs, represent a candidate cell source for cardiac cell therapy. However, it is not known what specific cell type or condition is the most appropriate for transplantation. This problem is exasperated by the lack of efficient and predictive strategies to screen the large numbers of parameters that may impact cell transplantation. We used a cardiac tissue model, engineered heart tissue (EHT), and quantitative molecular and electrophysiological analyses, to test transplantation conditions and specific cell populations for their potential to functionally integrate with the host tissue. In this study, we validated our analytical platform using contractile mouse neonatal CMs (nCMs) and noncontractile cardiac fibroblasts (cFBs), and screened for the integration potential of ESC-derived CMs and CPs (ESC-CMs and -CPs). Consistent with previous in vivo studies, cFB injection interfered with electrical signal propagation, whereas injected nCMs improved tissue function. Purified bioreactor-generated ESC-CMs exhibited a diminished capacity for electrophysiological integration; a result correlated with lower (compared with nCMs) connexin 43 expression. ESC-CPs, however, appeared able to appropriately mature and integrate into EHT, enhancing the amplitude of tissue contraction. Our results support the use of EHT as a model system to accelerate development of cardiac cell therapy strategies.

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“…A series of signal transduction systems have been implicated as essential coordinators of early cardiogenesis, including hedgehog (Hh), Wnt, bone morphogenetic protein (BMP) and platelet-derived growth factor receptor (PDGFR) signaling (Washington Smoak et al, 2005;Kwon et al, 2008;Hirata et al, 2007;van Wijk et al, 2007). In mammals, Hh signaling is induced by three different ligands, including Sonic hedgehog (Shh), which controls left-right asymmetry, digit patterning in the limbs and development of the lung and heart (Tsukui et al, 1999;Johnson et al, 1994;Bellusci et al, 1997;Washington Smoak et al, 2005).…”

Section: Introductionmentioning

confidence: 99%

The primary cilium coordinates early cardiogenesis and hedgehog signaling in cardiomyocyte differentiation

Clément

Kristensen

Møllgård

et al. 2009

Journal of Cell Science

100

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Defects in the assembly or function of primary cilia, which are sensory organelles, are tightly coupled to developmental defects and diseases in mammals. Here, we investigated the function of the primary cilium in regulating hedgehog signaling and early cardiogenesis. We report that the pluripotent P19.CL6 mouse stem cell line, which can differentiate into beating cardiomyocytes, forms primary cilia that contain essential components of the hedgehog pathway, including Smoothened, Patched-1 and Gli2. Knockdown of the primary cilium by Ift88 and Ift20 siRNA or treatment with cyclopamine, an inhibitor of Smoothened, blocks hedgehog signaling in P19.CL6 cells, as well as differentiation of the cells into beating cardiomyocytes. E11.5 embryos of the Ift88tm1Rpw (Ift88-null) mice, which form no cilia, have ventricular dilation, decreased myocardial trabeculation and abnormal outflow tract development. These data support the conclusion that cardiac primary cilia are crucial in early heart development, where they partly coordinate hedgehog signaling.

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Coexpression of platelet-derived growth factor receptor alpha and fetal liver kinase 1 enhances cardiogenic potential in embryonic stem cell differentiation in vitro

Cited by 30 publications

References 31 publications

GATA factors efficiently direct cardiac fate from embryonic stem cells

GATA factors efficiently direct cardiac fate from embryonic stem cells

Interrogating functional integration between injected pluripotent stem cell-derived cells and surrogate cardiac tissue

The primary cilium coordinates early cardiogenesis and hedgehog signaling in cardiomyocyte differentiation

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