Molecular characterization and functional properties of cardiomyocytes derived from human inducible pluripotent stem cells

The Sohnis Family Stem Cells Center, Technion - Israel Institute of Technology, Haifa, Israel.
Journal of Cellular and Molecular Medicine (Impact Factor: 4.01). 01/2011; 15(1):38-51. DOI: 10.1111/j.1582-4934.2009.00996.x
Source: PubMed


In view of the therapeutic potential of cardiomyocytes derived from induced pluripotent stem (iPS) cells (iPS-derived cardiomyocytes), in the present study we investigated in iPS-derived cardiomyocytes, the functional properties related to [Ca(2+) ](i) handling and contraction, the contribution of the sarcoplasmic reticulum (SR) Ca(2+) release to contraction and the b-adrenergic inotropic responsiveness. The two iPS clones investigated here were generated through infection of human foreskin fibroblasts (HFF) with retroviruses containing the four human genes: OCT4, Sox2, Klf4 and C-Myc. Our major findings showed that iPS-derived cardiomyocytes: (i) express cardiac specific RNA and proteins; (ii) exhibit negative force-frequency relations and mild (compared to adult) post-rest potentiation; (iii) respond to ryanodine and caffeine, albeit less than adult cardiomyocytes, and express the SR-Ca(2+) handling proteins ryanodine receptor and calsequestrin. Hence, this study demonstrates that in our cardiomyocytes clones differentiated from HFF-derived iPS, the functional properties related to excitation-contraction coupling, resemble in part those of adult cardiomyocytes.

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    • "Moving from dog CMs in an optical assay to rat or hiPSC-CMs in impedance introduces a new set of fundamental challenges which, if tolerable, may bring new opportunities. First, both the rat and hiPSC-CMs used in this study have an immature phenotype (Ma et al., 2011; Robertson et al., 2013) with the corresponding negative force–frequency relationship (Germanguz et al., 2011). This inverted relationship relative to in vivo raises the possibility that detection of inotropy could be severely limited or poorly translate. "

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    • "Our laboratory shows that NCX does not contribute to the calcium transient in ventricular hESC-CMs as indicated by the lack of effect of NCX inhibitors as well as its downregulation by short hairpin RNA [33]. Other calcium handling proteins normally present in adult CMs, including calsequestrin and phospholamban, have been shown to be absent in hESC-CMs [31,36], although there are reports that they are expressed in hESC-CMs [37,38]. The expression of sarco/endoplasmic reticulum Ca2+ ATPase (SERCA) pump in hESC-CMs is low and comparable to the levels in fetal CMs. "
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    ABSTRACT: Human pluripotent stem cells (hPSCs), including embryonic and induced pluripotent stem cells, are abundant sources of cardiomyocytes (CMs) for cell replacement therapy and other applications such as disease modeling, drug discovery and cardiotoxicity screening. However, hPSC-derived CMs display immature structural, electrophysiological, calcium-handling and metabolic properties. Here, we review various biological as well as physical and topographical cues that are known to associate with the development of native CMs in vivo to gain insights into the development of strategies for facilitated maturation of hPSC-CMs.
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    • "The cardiac differentiation ability of iPSCs and functionality of human iPSCs-derived cardiomyocytes have been characterized in previous reports [6], [7], [31], [32]. However, the only evaluation of functionality in derivatives of human pluripotent stem cells that contain genomic rearrangements in the TCR gene locus was in hepatic cells derived from human TiPSCs [33]. "
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    ABSTRACT: Induced pluripotent stem cells (iPSCs) have been proposed as novel cell sources for genetic disease models and revolutionary clinical therapies. Accordingly, human iPSC-derived cardiomyocytes are potential cell sources for cardiomyocyte transplantation therapy. We previously developed a novel generation method for human peripheral T cell-derived iPSCs (TiPSCs) that uses a minimally invasive approach to obtain patient cells. However, it remained unknown whether TiPSCs with genomic rearrangements in the T cell receptor (TCR) gene could differentiate into functional cardiomyocyte in vitro. To address this issue, we investigated the morphology, gene expression pattern, and electrophysiological properties of TiPSC-derived cardiomyocytes differentiated by floating culture. RT-PCR analysis and immunohistochemistry showed that the TiPSC-derived cardiomyocytes properly express cardiomyocyte markers and ion channels, and show the typical cardiomyocyte morphology. Multiple electrode arrays with application of ion channel inhibitors also revealed normal electrophysiological responses in the TiPSC-derived cardiomyocytes in terms of beating rate and the field potential waveform. In this report, we showed that TiPSCs successfully differentiated into cardiomyocytes with morphology, gene expression patterns, and electrophysiological features typical of native cardiomyocytes. TiPSCs-derived cardiomyocytes obtained from patients by a minimally invasive technique could therefore become disease models for understanding the mechanisms of cardiac disease and cell sources for revolutionary cardiomyocyte therapies.
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