Abstract 14104: Replacement of Infarct Myocardium by Large Scale-Expanded Human Induced Pluripotent Stem Cell-Derived Cardiac Cell-Sheet in a Porcine Chronic Myocardial Infarction Model

Circulation ◽  
2015 ◽  
Vol 132 (suppl_3) ◽  
Author(s):  
Keitaro Domae ◽  
Shigeru Miyagawa ◽  
Satsuki Fukushima ◽  
Atsuhiro Saito ◽  
Yukiko Imanishi ◽  
...  
Keyword(s):  
Functional Recovery ◽  
Large Scale ◽  
Induced Pluripotent Stem Cell ◽  
Cardiac Cells ◽  
Cell Group ◽  
Small Scale ◽  
Cardiac Cell ◽  
Cell Sheets ◽  
Induced Pluripotent ◽  
Human Ipsc

Introduction: It has been shown that transplanted induced pluripotent stem cell (iPSC)-derived cardiac cells in the myocardial infarction (MI) heart synchronously contract with native myocardium to mechanically contribute to functional recovery in rodent models. We herein hypothesized that large scale cardiac cell-sheets generated by human iPSCs may induce a greater functional recovery than small scale ones after transplantation in chronic MI heart. Methods: Bioreactor-based three-dimensional suspension culture system was used for generating large scale-expanded human iPSC-derived cardiomyocytes, of which cardiac troponin T positivity was constantly 75-85%. Scaffold-free cell-sheets containing several cell number (1.0х10^6, 10^7, 10^8) were transplanted over the cardiac surface in porcine chronic MI heart (n=5 each). Tacrolimus and prednisolone were daily given in all pigs against xeno-transplantation-inducing immune reaction. Results: Echocardiographically, left ventricular systolic and diastolic dimensions were significantly decreasing and ejection fraction was significantly increasing in the 10^8 cell group. In addition, global myocardial structure was better preserved in the 10^8 cell group with presence of the graft in the infarct area macroscopically (Figure). Moreover, there were significantly less accumulation of interstitial fibrosis in the infarct-remote area and greater vascular density and expression of VEGF, bFGF, and SDF-1 in the infarct-border area in the 10^8 cell group than the other groups at 3 months after the transplantation. Conclusions: Large scale human iPSC-derived cardiac cells were engrafted in the infarct myocardium, showing substantial functional recovery in a porcine chronic MI heart, indicating that artificial cell-based myocardial replacement therapy may be achieved. In contrast, small scale cardiac cells induced modest functional recovery, suggesting paracrine mechanisms of this treatment.

scholarly journals Screening for axon regeneration promoting compounds with human iPSC-derived motor neurons

2021 ◽  
Author(s):  
Tammy Szu-Yu Ho ◽  
J. Tabitha Hees ◽  
Zhuqiu Xu ◽  
Riki Kawaguchi ◽  
Natalia P Biscola ◽  
...  
Keyword(s):  
Nerve Injury ◽  
Motor Neurons ◽  
Large Scale ◽  
Axon Regeneration ◽  
Induced Pluripotent Stem Cell ◽  
Screening Strategy ◽  
Therapeutic Interventions ◽  
Induced Pluripotent ◽  
Human Ipsc

CNS neurons do not regenerate after injury, leading to permanent functional deficits. Although sensory and motor neuron axons do regrow after peripheral nerve injury, functional outcome is limited due to the incomplete and slow regrowth. The lack of human-relevant assays suitable for large-scale drug screens has limited neuro-repair therapy discovery. To address this we developed a phenotypic screening strategy using human induced pluripotent stem cell-derived motor neurons to identify axon-regeneration promoting compounds and targets. The screens involve both re-plating human motor neurons on chondroitin sulfate proteoglycans and measuring regeneration responses to laser axotomy in spot cultures, and from them we identified multiple hits that promote injured axon regrowth. The top hit blebbistatin, a non-muscle myosin II inhibitor, accelerated axon regeneration and functional recovery after sciatic nerve injury in vivo. Human injury in a dish assays are suitable, therefore, to screen for therapeutic interventions that can induce or accelerate axon regeneration.

scholarly journals SARS-CoV-2 infection of human iPSC-derived cardiac cells reflects cytopathic features in hearts of patients with COVID-19

2021 ◽  
pp. eabf7872
Author(s):  
Juan A. Perez-Bermejo ◽  
Serah Kang ◽  
Sarah J. Rockwood ◽  
Camille R. Simoneau ◽  
David A. Joy ◽  
...  
Keyword(s):  
Induced Pluripotent Stem Cell ◽  
Distinct Pattern ◽  
Cardiac Cells ◽  
Productive Infection ◽  
Heart Cells ◽  
Cardiac Damage ◽  
Autopsy Specimens ◽  
Induced Pluripotent ◽  
Human Ipsc

Although coronavirus disease 2019 (COVID-19) causes cardiac dysfunction in up to 25% of patients, its pathogenesis remains unclear. Exposure of human induced pluripotent stem cell (iPSC)-derived heart cells to severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2) revealed productive infection and robust transcriptomic and morphological signatures of damage, particularly in cardiomyocytes. Transcriptomic disruption of structural genes corroborates adverse morphologic features, which included a distinct pattern of myofibrillar fragmentation and nuclear disruption. Human autopsy specimens from patients with COVID-19 reflected similar alterations, particularly sarcomeric fragmentation. These striking cytopathic features in cardiomyocytes provide insights into SARS-CoV-2-induced cardiac damage, offer a platform for discovery of potential therapeutics, and raise concerns about the long-term consequences of COVID-19 in asymptomatic as well as severe cases.

Abstract 17227: Transplantation of Cardiac Cell Sheet Including Human Ips Cell-derived Cardiomyocytes and Vascular Cells to Infarcted Porcine Heart Restores Impaired Left Ventricular Global Function and Dyssynchrony

Circulation ◽  
2014 ◽  
Vol 130 (suppl_2) ◽  
Author(s):  
Masanosuke Ishigami ◽  
Hidetoshi Masumoto ◽  
Takayuki Aoki ◽  
Fumie Takai ◽  
Takeshi Ikuno ◽  
...  
Keyword(s):  
Systolic Function ◽  
Cell Sheet ◽  
Induced Pluripotent Stem Cell ◽  
Left Ventricular ◽  
Cardiac Cells ◽  
Border Region ◽  
Vascular Cells ◽  
Cardiac Cell ◽  
Cell Sheets ◽  
Ips Cell

BACKGROUNDS: Previously we reported beneficial effects of transplantation of all mouse ES cell-derived cardiac cell sheets composed of CMs and vascular cells (endothelial cells [ECs]/ vascular mural cells [MCs]) in a rat myocardial infarction (MI) model and importance of co-existence of vascular cells in the cell sheets (Stem Cells, 2012). Here we report a transplantation of human induced pluripotent stem cell (hiPSC)-derived cardiac cells to a porcine MI model. METHODS & RESULTS: We induced a mixture of CMs and vascular cells from hiPSCs with a 2-D serum-free method (modified from Uosaki, PLoS One, 2011). We generated cell sheets using 10cm-sized temperature-responsive culture dishes (UpCell; CellSeed, Tokyo, Japan). Self-pulsating cardiac cell sheets were approximately 3.5cm in diameter with 6.8х106±0.8 (n=5) of cells containing cTnT+-CMs (19.4±5.9%), VE-cadherin+-ECs (3.8±4.4%) and PDGFRβ+-MCs (67.2±7.8%). We induced MI in micromini-pigs (12-45 month old) by ameroid constriction of coronary artery and transplanted cell sheets (Tx) 2 weeks after MI induction (4 sheets / recipient) under immunosuppression. In Tx group, echocardiogram showed a significant improvement of systolic function of left ventricle (fractional shortening: 22.6±5.0 vs 39.7±8.7%, p<0.01, n=5). Ejection fraction evaluated by left ventriculogram improved significantly in Tx group (25.3±6.2% vs 39.8±4.2%, p<0.01, n=5). Speckle tracking echocardiogram showed significant increase of attenuated circumference strain in infarcted and border regions leading to restored dyssynchrony (anterior: 10.8±4.1 vs 20.6±3.5%, p<0.01, antero-lateral: 13±6.5% vs 24.8±7.6%, p<0.05, n=5, 2 weeks after Tx). Capillary density in the border region significantly elevated indicating angiogenic effect of the sheet transplantation (75.9±42.6/mm2 vs 137.4±44.8/mm2, p<0.001, n=5). CONCLUTION: HiPSC-derived cardiac cell sheets potentially ameliorate cardiac dysfunction of human-size infarct heart.

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

scholarly journals Contribution of two-pore K+ channels to cardiac ventricular action potential revealed using human iPSC-derived cardiomyocytes

2017 ◽  
Vol 312 (6) ◽  
pp. H1144-H1153 ◽  
Author(s):  
Sam Chai ◽  
Xiaoping Wan ◽  
Drew M. Nassal ◽  
Haiyan Liu ◽  
Christine S. Moravec ◽  
...  
Keyword(s):  
Action Potential ◽  
Expression Profile ◽  
Cardiac Electrophysiology ◽  
Human Heart ◽  
Induced Pluripotent Stem Cell ◽  
Heart Tissue ◽  
Induced Pluripotent ◽  
Interfering Rna ◽  
Human Ipsc ◽  
Physiological Systems

Two-pore K+ (K2p) channels have been described in modulating background conductance as leak channels in different physiological systems. In the heart, the expression of K2p channels is heterogeneous with equivocation regarding their functional role. Our objective was to determine the K2p expression profile and their physiological and pathophysiological contribution to cardiac electrophysiology. Induced pluripotent stem cells (iPSCs) generated from humans were differentiated into cardiomyocytes (iPSC-CMs). mRNA was isolated from these cells, commercial iPSC-CM (iCells), control human heart ventricular tissue (cHVT), and ischemic (iHF) and nonischemic heart failure tissues (niHF). We detected 10 K2p channels in the heart. Comparing quantitative PCR expression of K2p channels between human heart tissue and iPSC-CMs revealed K2p1.1, K2p2.1, K2p5.1, and K2p17.1 to be higher expressed in cHVT, whereas K2p3.1 and K2p13.1 were higher in iPSC-CMs. Notably, K2p17.1 was significantly lower in niHF tissues compared with cHVT. Action potential recordings in iCells after K2p small interfering RNA knockdown revealed prolongations in action potential depolarization at 90% repolarization for K2p2.1, K2p3.1, K2p6.1, and K2p17.1. Here, we report the expression level of 10 human K2p channels in iPSC-CMs and how they compared with cHVT. Importantly, our functional electrophysiological data in human iPSC-CMs revealed a prominent role in cardiac ventricular repolarization for four of these channels. Finally, we also identified K2p17.1 as significantly reduced in niHF tissues and K2p4.1 as reduced in niHF compared with iHF. Thus, we advance the notion that K2p channels are emerging as novel players in cardiac ventricular electrophysiology that could also be remodeled in cardiac pathology and therefore contribute to arrhythmias. NEW & NOTEWORTHY Two-pore K+ (K2p) channels are traditionally regarded as merely background leak channels in myriad physiological systems. Here, we describe the expression profile of K2p channels in human-induced pluripotent stem cell-derived cardiomyocytes and outline a salient role in cardiac repolarization and pathology for multiple K2p channels.

scholarly journals Contractile force measurement of human induced pluripotent stem cell-derived cardiac cell sheet-tissue

PLoS ONE ◽  
2018 ◽  
Vol 13 (5) ◽  
pp. e0198026 ◽  
Author(s):  
Daisuke Sasaki ◽  
Katsuhisa Matsuura ◽  
Hiroyoshi Seta ◽  
Yuji Haraguchi ◽  
Teruo Okano ◽  
...  
Keyword(s):  
Stem Cell ◽  
Pluripotent Stem Cell ◽  
Force Measurement ◽  
Cell Sheet ◽  
Induced Pluripotent Stem Cell ◽  
Contractile Force ◽  
Cardiac Cell ◽  
Induced Pluripotent

scholarly journals Robust Expression of Functional NMDA Receptors in Human Induced Pluripotent Stem Cell-Derived Neuronal Cultures Using an Accelerated Protocol

2021 ◽  
Vol 14 ◽  
Author(s):  
Jacob B. Ruden ◽  
Mrinalini Dixit ◽  
José C. Zepeda ◽  
Brad A. Grueter ◽  
Laura L. Dugan
Keyword(s):  
Nmda Receptors ◽  
Neural Progenitor Cells ◽  
Induced Pluripotent Stem Cell ◽  
Calcium Flux ◽  
Neuronal Cultures ◽  
Mature Neurons ◽  
Health And Disease ◽  
Nervous System Function ◽  
Induced Pluripotent ◽  
Human Ipsc

N-methyl-D-aspartate (NMDA) receptors are critical for higher-order nervous system function, but in previously published protocols to convert human induced pluripotent stem cells (iPSCs) to mature neurons, functional NMDA receptors (NMDARs) are often either not reported or take an extended time to develop. Here, we describe a protocol to convert human iPSC-derived neural progenitor cells (NPCs) to mature neurons in only 37 days. We demonstrate that the mature neurons express functional NMDARs exhibiting ligand-activated calcium flux, and we document the presence of NMDAR-mediated electrically evoked postsynaptic current. In addition to being more rapid than previous procedures, our protocol is straightforward, does not produce organoids which are difficult to image, and does not involve co-culture with rodent astrocytes. This could enhance our ability to study primate/human-specific aspects of NMDAR function and signaling in health and disease.

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