Transcriptional and Functional Profiling of Human Embryonic Stem Cell-Derived Cardiomyocytes

Department of Radiology, Stanford University School of Medicine, Stanford, California, United States of America.
PLoS ONE (Impact Factor: 3.23). 02/2008; 3(10):e3474. DOI: 10.1371/journal.pone.0003474
Source: PubMed


Human embryonic stem cells (hESCs) can serve as a potentially limitless source of cells that may enable regeneration of diseased tissue and organs. Here we investigate the use of human embryonic stem cell-derived cardiomyocytes (hESC-CMs) in promoting recovery from cardiac ischemia reperfusion injury in a mouse model. Using microarrays, we have described the hESC-CM transcriptome within the spectrum of changes that occur between undifferentiated hESCs and fetal heart cells. The hESC-CMs expressed cardiomyocyte genes at levels similar to those found in 20-week fetal heart cells, making this population a good source of potential replacement cells in vivo. Echocardiographic studies showed significant improvement in heart function by 8 weeks after transplantation. Finally, we demonstrate long-term engraftment of hESC-CMs by using molecular imaging to track cellular localization, survival, and proliferation in vivo. Taken together, global gene expression profiling of hESC differentiation enables a systems-based analysis of the biological processes, networks, and genes that drive hESC fate decisions, and studies such as this will serve as the foundation for future clinical applications of stem cell therapies.

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    • "Regarding myogenesis, in Drosophila, Twist has been shown to promote myogenesis. However, in mouse C2C12 myoblasts and in human embryonic stem cell (HESC)-derived embryoid bodies, Twist-1 is found to inhibit muscle cell differentiation (Hebrok et al., 1994; Rohwedel et al., 1995; Cao et al., 2008; Koutsoulidou et al., 2011a). Interestingly, overexpression of Twist-1 reverses the process of muscle cell differentiation (Hjiantoniou et al., 2008; Mastroyiannopoulos et al., 2013). "
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    ABSTRACT: Twist-1 is mostly expressed during development and has been previously shown to control myogenesis. Since its regulation in muscle has not been fully exploited, the aim of the project was to identify miRNAs in muscle which regulate Twist-1. miR-206, one of the most important myomiRs, was identified as a possible candidate for Twist-1 mRNA. Luciferase assays and transfections in human foetal myoblasts showed that Twist-1 is a direct target for miR-206 and through this pathway muscle cell differentiation is promoted. We next investigated whether MyoD, a major myogenic transcription factor regulates Twist-1, since it is known that MyoD induces miR-206 gene expression. We found that forced MyoD expression induces miR-206 up-regulation and Twist-1 down-regulation through miR-206 promoter binding, followed by increase in muscle cell differentiation. Finally, experiments were performed in muscle cells from patients with congenital Myotonic Dystrophy type 1 which fail to differentiate to myotubes. MyoD overexpression inhibited Twist-1 through miR-206 induction, followed by an increase in muscle cell differentiation. These results reveal a novel mechanism of myogenesis which might also play an important role in muscle disease. © 2015. Published by The Company of Biologists Ltd.
    No preview · Article · Aug 2015 · Journal of Cell Science
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    • "Monolayer culture High purity hESC-and hiPSC-derived CMs Cao et al. 2008 [59] Suspension EB Expression level of genes encoding structural and force-generating proteins was comparable to fetal heart CMs Caspi et al. 2009 [74] Suspension EB Presence of functional gap junctions Chan et al. 2013 [102] Suspension EB "
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    ABSTRACT: Human pluripotent stem cell-derived cardiomyocytes (hPSC-CMs) are the most promising source of cardiomyocytes (CMs) for experimental and clinical applications, but their use is largely limited by a structurally and functionally immature phenotype that most closely resembles embryonic or fetal heart cells. The application of physical stimuli to influence hPSC-CMs through mechanical and bioelectrical transduction offers a powerful strategy for promoting more developmentally mature CMs. Here we summarize the major events associated with in vivo heart maturation and structural development. We then review the developmental state of in vitro derived hPSC-CMs, while focusing on physical (electrical and mechanical) stimuli and contributory (metabolic and hypertrophic) factors that are actively involved in structural and functional adaptations of hPSC-CMs. Finally, we highlight areas for possible future investigation that should provide a better understanding of how physical stimuli may promote in vitro development and lead to mechanistic insights. Advances in the use of physical stimuli to promote developmental maturation will be required to overcome current limitations and significantly advance research of hPSC-CMs for cardiac disease modeling, in vitro drug screening, cardiotoxicity analysis and therapeutic applications.
    Full-text · Article · Oct 2014 · Stem Cell Research & Therapy
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    • "expressed gene in pluripotent hES cells vs. their differentiated counterparts (Cao et al., 2008). In addition, a global phosphoproteomic study of hES cells identified Lck as a signaling protein with more abundant phosphorylation site identifications in pluripotent hES cells compared to differentiated hES cell derivatives (Brill et al., 2009). "
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    ABSTRACT: Embryonic stem (ES) cells are characterized by pluripotency, defined as the developmental potential to generate cell lineages derived from all three primary germ layers. In the past decade, great progress has been made on the cell culture conditions, transcription factor programs and intracellular signaling pathways that control both murine and human ES cell fates. ES cells of mouse vs. human origin have distinct culture conditions, responding to some tyrosine kinase signaling pathways in opposite ways. Previous work has implicated the Src family of non-receptor protein-tyrosine kinases in mouse ES cell self-renewal and differentiation. Seven members of the Src kinase family are expressed in mouse ES cells, and individual family members appear to play distinct roles in regulating their developmental fate. Both Hck and c-Yes are important in self-renewal, while c-Src activity alone is sufficient to induce differentiation. While these findings implicate Src-family kinase signaling in mouse ES cell renewal and differentiation, the role of this kinase family in human ES cells is largely unknown. Here, we explored Src-family kinase expression patterns and signaling in human ES cells during self-renewal and differentiation. Of the eleven Src-related kinases in the human genome, Fyn, c-Yes, c-Src, Lyn, Lck and Hck were expressed in H1, H7 and H9 hES cells, while Fgr, Blk, Srm, Brk, and Frk transcripts were not detected. Of these, c-Yes, Lyn, and Hck transcript levels remained constant in self-renewing human ES cells vs. differentiated EBs, while c-Src and Fyn showed a modest increase in expression as a function of differentiation. In contrast, Lck expression levels dropped dramatically as a function of EB differentiation. To assess the role of overall Src-family kinase activity in human ES cell differentiation, cultures were treated with inhibitors specific for the Src kinase family. Remarkably, human ES cells maintained in the presence of the potent Src-family kinase inhibitor A-419259 retained the morphology of domed, pluripotent colonies and continued to express the self-renewal marker TRA-1-60 despite culture under differentiation conditions. Taken together, these observations support a role for Src-family kinase signaling in the regulation of human ES cell fate, and suggest that the activities of individual Src-family members are required for initiation of the differentiation program.
    Full-text · Article · Sep 2014 · Stem Cell Research
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