Abstract 16007: Functional Comparison of Human Embryonic Stem Cell- versus Human Induced Pluripotent Stem Cell-Derived Cardiomyocytes in Rat Ischemic Myocardial Infarction Model

Circulation ◽  
2014 ◽  
Vol 130 (suppl_2) ◽  
Author(s):  
Wenyi Chen ◽  
Johannes Riegler ◽  
Elena Matsa ◽  
Qi Shen ◽  
Haodi Wu ◽  
...  
Keyword(s):  
Myocardial Infarction ◽  
Stem Cell ◽  
Cardiac Function ◽  
Embryonic Stem Cell ◽  
Pluripotent Stem Cell ◽  
Human Embryonic Stem Cell ◽  
Embryonic Stem ◽  
Induced Pluripotent Stem Cell ◽  
Left Ventricular ◽  
Induced Pluripotent

Introduction: Both human embryonic stem cell-derived cardiomyocytes (ESC-CMs) and human induced pluripotent stem cell-derived cardiomyocytes (iPSC-CMs) can serve as an unlimited cell source for cardiac regenerative therapy. However, the functional equivalency of both approaches has not been previously reported. Here we performed head-to-head comparison on the beneficial effects of ESC-CM and iPSC-CMs in restoring cardiac function in a rat myocardial infarction (MI) model. Methods & Results: Human ESCs and iPSCs were differentiated into cardiomyocytes using small molecules. FACS analysis confirmed ~85% and ~83% of cells differentiated from ESCs and iPSCs, respectively, were positive for cardiac troponin T, and immunofluorescence staining demonstrated that ESC-CMs and iPSC-CMs have striated sarcomeric structure (Figure A-B). Both ESC-CMs and iPSC-CMs displayed similar maturity for calcium handling (transient amplitude: ΔF/F 0 = 3.8±0.3; time to peak: ~200 ms; 50% transient duration: ~400 ms). qRT-PCR showed that ESC-CMs and iPSC-CMs expressed CASQ2, GJA5, KCNJ2, KCNJ5, MYH6, MYH7, and SCN5A at comparable levels to human fetal heart tissue. Next, ESC-CMs and iPSC-CMs were injected into the left ventricular free wall of infarcted hearts (adult nude rats; n=14, 10, respectively). Cardiac function was assessed by MRI one month post cell injection and the hearts were harvested and stained for human cardiac markers. Both ESC-CMs and iPSC-CMs could engraft in ischemic rat hearts (Figure C). Comprehensive functional analysis with small animal magnetic resonance imaging (MRI), echocardiography, and pressure-volume loop analysis are underway. Conclusion: We set out to perform head to head comparison for the first time that iPSC-CMs may facilitate cardiac repair at comparable levels to ESC-CMs. Unlike allogeneic ESC-CM therapy, autologous iPSC-CMs could be used to overcome immune rejection for cardiac cell transplantation in the future.

scholarly journals A Novel Purification Method of Murine Embryonic Stem Cell– and Human-Induced Pluripotent Stem Cell–Derived Cardiomyocytes by Simple Manual Dissociation

2012 ◽  
Vol 17 (5) ◽  
pp. 683-691 ◽  
Author(s):  
Tadahiro Shinozawa ◽  
Hatsue Furukawa ◽  
Eimei Sato ◽  
Kenji Takami
Keyword(s):  
Stem Cells ◽  
Stem Cell ◽  
Embryonic Stem Cell ◽  
Pluripotent Stem Cell ◽  
Embryonic Stem ◽  
Induced Pluripotent Stem Cell ◽  
Drug Induced ◽  
Thin Glass ◽  
Murine Embryonic Stem Cell ◽  
Induced Pluripotent

Cardiomyocytes derived from embryonic stem cells (ES-CMs) and induced pluripotent stem cells (iPS-CMs) are useful for toxicity and pharmacology screening. In the present study, we found that cardiomyocyte-rich beating cell clusters (CCs) emerged from murine embryonic stem cell (mESC)–derived beating EBs and from human-induced pluripotent stem cell (hiPSC)–derived beating EBs dissociated by gentle pipetting with a thin glass pipette. The percentage of cardiac troponin T (cTnT)–positive cells in the beating CCs obtained from mESC-derived and hiPSC-derived beating EBs was higher (81.5% and 91.6%, respectively) than in beating-undissociated EBs (13.7% and 67.1%, respectively). For mESCs, the yield of cTnT-positive cells from beating CCs was estimated to be 1.6 times higher than that of beating EBs. The bromodeoxyuridine labeling index of mouse ES-CMs and human iPS-CMs in beating CCs was 1.5- and 3.2-fold, respectively, greater than those in beating EBs. To investigate the utility of the cells in toxicity assessment, we showed that doxorubicin, a cardiotoxic drug, induced myofilament disruption in cardiomyocytes isolated by this method. This simple method enables preparation of mouse ES-CMs and human iPS-CMs with better proliferative activity than beating EBs not dissociated by pipetting, and the cardiomyocytes are useful for drug-induced myocardial toxicity testing.

scholarly journals Sacubitril/Valsartan Improves Cardiac Function and Decreases Myocardial Fibrosis Via Downregulation of Exosomal miR‐181a in a Rodent Chronic Myocardial Infarction Model

2020 ◽  
Vol 9 (13) ◽  
Author(s):  
Evgeniya Vaskova ◽  
Gentaro Ikeda ◽  
Yuko Tada ◽  
Christine Wahlquist ◽  
Marc Mercola ◽  
...  
Keyword(s):  
Myocardial Infarction ◽  
Stem Cell ◽  
Cardiac Function ◽  
Pluripotent Stem Cell ◽  
Myocardial Injury ◽  
Induced Pluripotent Stem Cell ◽  
Injury Model ◽  
Induced Pluripotent

Background Exosomes are small extracellular vesicles that function as intercellular messengers and effectors. Exosomal cargo contains regulatory small molecules, including mi RNA s, mRNA s, lnc RNA s, and small peptides that can be modulated by different pathological stimuli to the cells. One of the main mechanisms of action of drug therapy may be the altered production and/or content of the exosomes. Methods and Results We studied the effects on exosome production and content by neprilysin inhibitor/angiotensin receptor blockers, sacubitril/valsartan and valsartan alone, using human‐induced pluripotent stem cell‐derived cardiomyocytes under normoxic and hypoxic injury model in vitro , and assessed for physiologic correlation using an ischemic myocardial injury rodent model in vivo. We demonstrated that the treatment with sacubitril/valsartan and valsartan alone resulted in the increased production of exosomes by induced pluripotent stem cell‐derived cardiomyocytes in vitro in both conditions as well as in the rat plasma in vivo. Next‐generation sequencing of these exosomes exhibited downregulation of the expression of rno‐miR‐181a in the sacubitril/valsartan treatment group. In vivo studies employing chronic rodent myocardial injury model demonstrated that miR‐181a antagomir has a beneficial effect on cardiac function. Subsequently, immunohistochemical and molecular studies suggested that the downregulation of miR‐181a resulted in the attenuation of myocardial fibrosis and hypertrophy, restoring the injured rodent heart after myocardial infarction. Conclusions We demonstrate that an additional mechanism of action of the pleiotropic effects of sacubitril/valsartan may be mediated by the modulation of the mi RNA expression level in the exosome payload.

Abstract 17089: Cyclin D2 Overexpression Enhances the Potency of Human Induced-Pluripotent Stem Cell-Derived Cardiomyocytes for Cardiac Repair in a Porcine Model of Myocardial Infarction

Circulation ◽  
2018 ◽  
Vol 138 (Suppl_1) ◽  
Author(s):  
Meng Zhao ◽  
Yawen Tang ◽  
Wuqiang Zhu ◽  
Jianyi Zhang
Keyword(s):  
Myocardial Infarction ◽  
Cell Cycle ◽  
Stem Cell ◽  
Cardiac Function ◽  
Pluripotent Stem Cell ◽  
Myocardial Injury ◽  
Induced Pluripotent Stem Cell ◽  
Large Animal ◽  
Cyclin D2 ◽  
Induced Pluripotent

Introduction: Stem-cell-based therapies provide promise for the treatment of ischemic myocardial injury, but the number of cells that remain engrafted at the site of administration is exceptionally low, which is believed to limit the effectiveness of treatment. In this study, we investigated whether the number of engrafted human induced-pluripotent stem-cell-derived cardiomyocytes (hiPSC-CMs) can be increased via overexpression of the cell-cycle activator cyclin D2 (CCND2) and if so, whether this increase is accompanied by improvements in myocardial recovery. Experiments were conducted in a large-animal (swine) model of myocardial injury. Methods and results: CCND2-overexpressing hiPSC-CMs (CCND2 OE CMs) were constructed via lentiviral transfection; then, myocardial infarction (MI) was surgically induced in swine, and the animals were randomly assigned to treatment with CCND2 OE CMs (i.e., the MI+CCND2 OE CM group), wild-type hiPSC-CMs (the MI+CCND2 WT CM group), or neither experimental treatment (the MI group); the cells were injected into five sites (6х10 6 cells/site) surrounding the infarct. Four weeks after MI, measurements of the heart-weight to bodyweight ratio (HW/BW), cardiac function (determined via magnetic resonance imaging), and vascularity (CD31 staining) were significantly better in MI+CCND2 OE CM animals than in the CCND2 WT CM group. HW/BW was also significantly lower in both cell-treatment groups than in MI animals, and CCND2 overexpression was associated with increases in both hiPSC-CM engraftment (mRNA levels) and cell-cycle activity (ki67 and phosphorylated histone 3 levels). Conclusion: Experiments in a large-animal MI model indicated that CCND2 overexpression in transplanted hiPSC-CMs was associated with increases in cell-cycle activity and the number of engrafted cells, as well as improvements in cardiac function, hypertrophy, and vascularity.

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