Abstract 14377: Therapeutic Potential of Clinical Grade Human Induced Pluripotent Stem Cell-derived Cardiac Tissues for a Rat Myocardial Infarction Model: A Pre-clinical Study for a Cell Transplantation Therapy

Circulation ◽  
2020 ◽  
Vol 142 (Suppl_3) ◽  
Author(s):  
Hiroaki Osada ◽  
Hidetoshi Masumoto ◽  
Masahide Kawatou ◽  
Tadashi Ikeda ◽  
Yasuhiko Tabata ◽  
...  
Keyword(s):  
Myocardial Infarction ◽  
Stem Cell ◽  
Pluripotent Stem Cell ◽  
Induced Pluripotent Stem Cell ◽  
Left Ventricular ◽  
Clinical Grade ◽  
Cell Sheets ◽  
Cardiac Tissues ◽  
Cell Lineages ◽  
Induced Pluripotent

Introduction: Transplantation of three-dimensional bioengineered cardiac tissues composed of pluripotent stem cell-derived cardiovascular cell lineages is reported to hold potential for functional recovery on pre-clinical studies. We aimed to evaluate therapeutic and myocardial regenerative potential of clinical grade human induced pluripotent stem cell (hiPSC)-derived cardiac tissues (HiCTs) on a rat myocardial infarction (MI) model. Methods: Clinical grade hiPSC lines established from a healthy volunteer were simultaneously differentiated into cardiovascular cell lineages. We seeded the cells on temperature responsive culture dishes to form cell sheets. HiCTs were generated by stacking 5 cell sheets with insertion of gelatin hydrogel microspheres (GHMs) to promote oxygen and nutrition supply and transplanted onto an athymic rat MI model (n=6). Echocardiography and histological analysis at 12 weeks after surgery were conducted and compared to those in animals with sham surgery (n=9) and with cell sheet stacks without GHMs [GHM(-), n=6]. Some of HiCT-transplanted rats were subjected to tissue clearing and two-photon excitation microscopy (TPEM) to assess graft vascularization. Results: Flow cytometry revealed cellular components after differentiation as follows: 52.0±1.4% of cardiomyocytes (cardiac isoform of troponin-T + :cTnT), 9.9±0.7% of vascular endothelial cells (VE-cadherin + ) and 14.1±1.8% of vascular mural cells (PDGFRβ + ). Echocardiography revealed significantly lower left ventricular end diastolic volume (LVEDV) and higher left ventricular ejection fraction (LVEF) in HiCT group [sham vs GHM(-) vs HiCT: LVEDV; 1.5±0.1 vs 1.3±0.05 vs 0.9±0.03 mL, p<0.0001 / LVEF; 59.7±2.2 vs 67.4±0.6 vs 82.7±0.9 %, p<0.0001]. HiCT group showed significantly larger engraftment [GHM(-) vs HiCT; 12 week; 0.1 ± 0.1 vs 1.8 ± 0.6 mm 2 ; p<0.05]. Engrafted graft tissues were composed of cTnT / α-Actinin-positive cardiomyocytes which exhibited obvious striated structure. TPEM revealed host to graft vascular connection at 2 weeks after HiCT transplantation. Conclusions: HiCTs derived from clinical grade hiPSC potentially serve as a stem cell-derived cellular product in cardiac regenerative therapy for foreseeable clinical applications.

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 Therapeutic potential of clinical-grade human induced pluripotent stem cell-derived cardiac tissues

JTCVS Open ◽  
2021 ◽  
Author(s):  
Hiroaki Osada ◽  
Masahide Kawatou ◽  
Daiki Fujita ◽  
Yasuhiko Tabata ◽  
Kenji Minatoya ◽  
...  
Keyword(s):  
Stem Cell ◽  
Pluripotent Stem Cell ◽  
Therapeutic Potential ◽  
Induced Pluripotent Stem Cell ◽  
Clinical Grade ◽  
Cardiac Tissues ◽  
Induced Pluripotent

Transplantation of clinical-grade human induced pluripotent stem cell derived cardiac tissues contributes to functional recovery in a rat myocardial infarction model

2020 ◽  
Vol 41 (Supplement_2) ◽  
Author(s):  
H Osada ◽  
H Masumoto ◽  
M Kawatou ◽  
T Ikeda ◽  
Y Tabata ◽  
...  
Keyword(s):  
Functional Recovery ◽  
Left Ventricular ◽  
Funding Source ◽  
Clinical Grade ◽  
Cell Sheets ◽  
Cardiac Tissues ◽  
Cell Lineages ◽  
Cell Source ◽  
Induced Pluripotent ◽  
Human Ipsc

Abstract Background Clinical-grade human induced pluripotent stem cells (iPSCs) established from a healthy volunteer are currently being considered as a quality controlled cell source for regenerative therapy. Transplantation of three-dimensional bioengineered cardiac tissues composed of human iPSC-derived cardiovascular cell lineages is reported to hold potential for cardiac functional recovery. Purpose The aims of this study were to evaluate tissue conformation and cellular viability of human iPSC-derived cardiac tissues (HiCTs) generated from clinical-grade cells and to validate functional efficacy of HiCT transplantation. Methods Clinical-grade human iPSC lines were simultaneously differentiated into cardiovascular cell lineages by a high-density monolayer culture. The differentiation efficacy was analyzed by flow cytometry. We seeded the cells on temperature responsive culture dishes to form cell sheets. HiCTs are generated by stacking 5 cell sheets with insertion of gelatin hydrogel microspheres (GHMs) between each sheet to promote oxygen and nutrition supply. Characteristics of the HiCTs are histologically and immunohistochemically evaluated. The HiCTs were transplanted onto an athymic nude rat myocardial infarction (MI) model. Cardiac function was evaluated by echocardiography and cardiac magnetic resonance imaging (MRI) until 4 weeks after surgery, and compared to those in animals with sham operation and with cell sheet stacks without GHMs [GHM(−)]. Results Flow cytometry at differentiation day15 revealed cellular components as follows: 52.5±1.4% of cardiomyocytes (cardiac isoform of troponin-T+), 9.8±0.7% of vascular endothelial cells (VE-cadherin+), 14.8±1.8% of vascular mural cells (PDGFRβ+) and 0.2±0.1% of undifferentiated cells (TRA-1-60+). HiCTs were significantly thicker [GHM(−) vs HiCT: 357.3±81.5 vs 723.0±84.0μm, p&lt;0.05], composed of higher area of cardiomyocytes (27.7±7.9 vs 71.9±15.5mm2, p&lt;0.05) and endothelial cells (CD31+) (1.6±0.7 vs 9.2±1.5mm2, p&lt;0.05), free from hypoxia (HIF-1α+) (3.1±0.1 vs 0.8±0.2%, p&lt;0.05) and cell death (TUNEL+) (3.2±0.1 vs 1.4±0.3%, p&lt;0.05) after 7 days of in vitro culture. Echocardiography revealed significantly lower left ventricular end diastolic volume (LVEDV) and higher left ventricular ejection fraction (LVEF) in HiCT group [sham (n=27) vs GHM(−) (n=12) vs HiCT (n=12): LVEDV; 1.4±0.1 vs 1.3±0.1 vs 0.9±0.1mL, p&lt;0.0001/LVEF; 55.3±1.1 vs 58.2±2.3 vs 78.2±1.5%, p&lt;0.0001]. Cardiac MRI showed lower LVEDV and higher LVEF as well [sham (n=8) vs GHM(−) (n=6) vs HiCT (n=6): LVEDV; 0.7±0.03 vs 0.7±0.03 vs 0.6±0.02mL, p&lt;0.01 / LVEF; 39.2±2.1 vs 43.8±1.4 vs 54.0±2.8%, p&lt;0.001]. Conclusions We conclude that HiCTs generated from clinical-grade cells hold sufficient viability and tissue conformation suitable for functional recovery validated by a rat MI model. Clinical-grade human iPSCs potentially serve as a reasonable cell source for stem cell-derived product transplantation therapy with foreseeable clinical applications. Funding Acknowledgement Type of funding source: Other. Main funding source(s): Japan Agency for Medical Research and Development (AMED), Invited Research Project of Institute for Advancement of Clinical Transnational Science, Kyoto University Hospital

Abstract 205: Feasibility and Testing of a Human Induced Pluripotent Stem Cell Derived Cardiac Patch in a Pre-clinical Swine Model of CHF

2016 ◽  
Vol 119 (suppl_1) ◽  
Author(s):  
Jordan J Lancaster ◽  
Ike Chinyere ◽  
Bin Na Kim ◽  
Sherry Daugherty ◽  
Samuel Kim ◽  
...  
Keyword(s):  
Quality Of Life ◽  
Stem Cell ◽  
Pluripotent Stem Cell ◽  
Induced Pluripotent Stem Cell ◽  
Left Ventricular ◽  
Dermal Fibroblasts ◽  
Swine Model ◽  
Induced Pluripotent ◽  
Cardiac Patch

Introduction: Previously we have demonstrated that a tissue engineered heart patch comprised of human induced pluripotent stem cell derived cardiomyocytes (hiPSC-CMs) and fibroblasts improves both left ventricular (LV) systolic and diastolic function in a rat model of CHF. In this study we tested the feasibility of upscaling cardiac patch size and surgical deployment in a swine model of CHF to test clinical utility. Methods: Four male Gottingen mini swine 20-25kg and three domestic swine 50-60kg were infarcted using percutaneous methods. Embolizing coils were deployed via catheter distal to the first diagonal branch of the left anterior descending (LAD) coronary artery and animals recovered for 4 weeks. Cardiac patches engineered with bio absorbable polygalactin-910 knitted mesh, dermal fibroblasts and hiPSC-CMs were cultured and implanted on the infarcted epicardium 4 weeks after MI. Cardiac magnetic resonance imaging was performed at baseline, 4 and 8 weeks post MI. All swine were implanted with continuous event recorders to acquire surface electrocardiogram during the entire study. In addition quality of life and functional capacity were assessed through video monitoring and treadmill exertion testing respectively. Infarct size was determined through 2,3,5-triphenyltetrazolium chloride staining. Results: LAD occlusion resulted in a significant (P<0.05) decrease EF (15%), and increase in EDV (59%) and ESV (100%). Average TIMI score decreased from 3.0±0 at time of MI to 1.5±0.6 4wks post MI. Cardiac patches were upsized to 6cm diameter for application in the swine. Patches displayed synchronous and spontaneous contractions within 48hrs. The 6cm patches, when implanted effectively covered the infarcted region bridging viable myocardium. Surgical handling and epicardial deployment was successfully accomplished via median sternotomy. The patches were robust in nature and could be deployed via a minimally invasive robotic procedure. No adverse arrhythmic activity was observed. Implantation of the cardiac patch restored activity levels (quality of life) of patch treated swine vs CHF controls. Conclusion: Our hiPSC-CM cardiac patch can be constructed in a clinical size, easily handled and implanted on the epicardium of the infarcted heart.

scholarly journals N-cadherin overexpression enhances the reparative potency of human-induced pluripotent stem cell-derived cardiac myocytes in infarcted mouse hearts

2019 ◽  
Vol 116 (3) ◽  
pp. 671-685 ◽  
Author(s):  
Xi Lou ◽  
Meng Zhao ◽  
Chengming Fan ◽  
Vladimir G Fast ◽  
Mani T Valarmathi ◽  
...  
Keyword(s):  
Stem Cell ◽  
Infarct Size ◽  
Cardiac Myocytes ◽  
Pluripotent Stem Cell ◽  
Myogenic Differentiation ◽  
Functional Integration ◽  
Induced Pluripotent Stem Cell ◽  
Left Ventricular ◽  
Cardiac Functions ◽  
Induced Pluripotent

Abstract Aims In regenerative medicine, cellular cardiomyoplasty is one of the promising options for treating myocardial infarction (MI); however, the efficacy of such treatment has shown to be limited due to poor survival and/or functional integration of implanted cells. Within the heart, the adhesion between cardiac myocytes (CMs) is mediated by N-cadherin (CDH2) and is critical for the heart to function as an electromechanical syncytium. In this study, we have investigated whether the reparative potency of human-induced pluripotent stem cell-derived cardiac myocytes (hiPSC-CMs) can be enhanced through CDH2 overexpression. Methods and results CDH2-hiPSC-CMs and control wild-type (WT)-hiPSC-CMs were cultured in myogenic differentiation medium for 28 days. Using a mouse MI model, the cell survival/engraftment rate, infarct size, and cardiac functions were evaluated post-MI, at Day 7 or Day 28. In vitro, conduction velocities were significantly greater in CDH2-hiPSC-CMs than in WT-hiPSC-CMs. While, in vivo, measurements of cardiac functions: left ventricular (LV) ejection fraction, reduction in infarct size, and the cell engraftment rate were significantly higher in CDH2-hiPSC-CMs treated MI group than in WT-hiPSC-CMs treated MI group. Mechanistically, paracrine activation of ERK signal transduction pathway by CDH2-hiPSC-CMs, significantly induced neo-vasculogenesis, resulting in a higher survival of implanted cells. Conclusion Collectively, these data suggest that CDH2 overexpression enhances not only the survival/engraftment of cultured CDH2-hiPSC-CMs, but also the functional integration of these cells, consequently, the augmentation of the reparative properties of implanted CDH2-hiPSC-CMs in the failing hearts.

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