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.

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.

Abstract 020: Cell-sheet-based Bioengineered Human Cardiac Tissue Using Pluripotent Stem Cells For Heart Repair And Disease Models

2013 ◽  
Vol 113 (suppl_1) ◽  
Author(s):  
Katsuhisa Matsuura ◽  
Tatsuya Shimizu ◽  
Nobuhisa Hagiwara ◽  
Teruo Okano
Keyword(s):  
Cardiac Tissue ◽  
Subcutaneous Tissue ◽  
Cell Sheet ◽  
Three Dimensional ◽  
Embryoid Bodies ◽  
Cardiac Differentiation ◽  
Cardiac Cell ◽  
Cell Sheets ◽  
High Efficient ◽  
Human Cardiac Tissue

We have developed an original scaffold-free tissue engineering approach, “cell sheet engineering”, and this technology has been already applied to regenerative medicine of various organs including heart. As the bioengineered three-dimensional cardiac tissue is expected to not only function for repairing the broad injured heart but also to be the practicable heart tissue models, we have developed the cell sheet-based perfusable bioengineered three-dimensional cardiac tissue. Recently we have also developed the unique suspension cultivation system for the high-efficient cardiac differentiation of human iPS cells. Fourteen-day culture with the serial treatments of suitable growth factors and a small compound in this stirring system with the suitable dissolved oxygen concentration produced robust embryoid bodies that showed the spontaneous beating and were mainly composed of cardiomyocytes (~80%). When these differentiated cells were cultured on temperature-responsive culture dishes after the enzymatic dissociation, the spontaneous and synchronous beating was observed accompanied with the intracellular calcium influx all over the area even after cell were detached from culture dishes as cell sheets by lowering the culture temperature. The cardiac cell sheets were mainly composed of cardiomyocytes (~80%) and partially mural cells (~20%). Furthermore, extracellular action potential propagation was observed between cell sheets when two cardiac cell sheets were partially overlaid, and this propagation was inhibited by the treatment with some anti-arrhythmic drugs. When the triple layered cardiac tissue was transplanted onto the subcutaneous tissue of nude rats, the spontaneous pulsation was observed over 2 months and engrafted cardiomyocytes were vascularized with the host tissue-derived endothelial cells. These findings suggest that cardiac cell sheets formed by hiPSC-derived cardiomyocytes might have sufficient properties for the creation of thickened cardiac tissue. Now we are developing the vascularized thickened human cardiac tissue by the repeated layering of cardiac cell sheets on the artificial vascular bed in vitro.

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 Stem cell-derived cell sheet transplantation for heart tissue repair in myocardial infarction

2020 ◽  
Vol 11 (1) ◽  
Author(s):  
Rui Guo ◽  
Masatoshi Morimatsu ◽  
Tian Feng ◽  
Feng Lan ◽  
Dehua Chang ◽  
...  
Keyword(s):  
Myocardial Infarction ◽  
Stem Cell ◽  
Tissue Repair ◽  
Cardiac Tissue ◽  
Cell Sheet ◽  
Clinical Manifestations ◽  
Induced Pluripotent Stem Cell ◽  
Heart Tissue ◽  
Therapeutic Effects ◽  
Cell Sheets

AbstractStem cell-derived sheet engineering has been developed as the next-generation treatment for myocardial infarction (MI) and offers attractive advantages in comparison with direct stem cell transplantation and scaffold tissue engineering. Furthermore, induced pluripotent stem cell-derived cell sheets have been indicated to possess higher potential for MI therapy than other stem cell-derived sheets because of their capacity to form vascularized networks for fabricating thickened human cardiac tissue and their long-term therapeutic effects after transplantation in MI. To date, stem cell sheet transplantation has exhibited a dramatic role in attenuating cardiac dysfunction and improving clinical manifestations of heart failure in MI. In this review, we retrospectively summarized the current applications and strategy of stem cell-derived cell sheet technology for heart tissue repair in MI.

Abstract 153: Creation of Cell Sheet-Based Bioengineered Cardiac Tissue Using Pluripotent Stem Cell-Derived Cells

2012 ◽  
Vol 111 (suppl_1) ◽  
Author(s):  
Katsuhisa Matsuura ◽  
Shinako Aoki ◽  
Yuji Haraguchi ◽  
Tatsuya Shimizu ◽  
Nobuhisa Hagiwara ◽  
...  
Keyword(s):  
Stem Cell ◽  
Pluripotent Stem Cell ◽  
Cardiac Tissue ◽  
Es Cells ◽  
Cell Sheet ◽  
Ips Cells ◽  
Cardiac Differentiation ◽  
Cardiac Cell ◽  
Cell Sheets ◽  
Mouse Es Cells

Recent evidences have suggested that current cardiac cell therapy contributes to the improved cardiac function through mainly the paracrine effects. Thereby the bioengineered functional heart tissue is expected to function for repairing the broad injured heart. We have developed the cell sheet-based bioengineered vascularized cardiac tissue, however the system to collect the enough amount of cells from ES/iPS cells and the function of ES-derived cardiac tissue remain elusive. Recently we have established the cultivation system with the suitable conditions for expansion and cardiac differentiation of mouse ES cells and human iPS cells via embryoid body formation using three-dimensional bioreactor with the continuous perfusion system. For the cardiac differentiation experiments, we used several mouse ES cells that express EGFP or neomycin resistant gene under the control of αMHC promoter. At 10 days of differentiation, mouse ES cells increased up to 300 times (6.0×10 6 cells/mL). After the further 8 days of cultivation with the purification step, we collected around 5.0×10 8 cells in the 1L bioreactor culture and 99% of cells were positive for myosin heavy chain. The co-culture of ES-derived cardiomyocytes with the appropriate number of primary cultured fibroblasts on the temperature-responsive culture dishes enabled to form the cardiac cell sheets. Furthermore, when ES-derived cardiomyocytes were co-cultured with ES-derived endothelial cells, robust endothelial cell network was observed in the cardiac cell sheets. Mouse ES- or human iPS-derived cardiomyocytes in cell sheets beat spontaneously and synchronously and connexin43 was expressed at the edge of the adjacent cardiomyocytes. Furthermore the action potential propagation was observed between ES/iPS-derived cardiac cell sheets. These findings suggest that pluripotent stem cell-derived cardiomyocytes and endothelial cells might be useful for creating cell-sheet-based functional cardiac tissue and the layered stem cell-derived cardiac tissue might promote not only the cardiac regenerative medicine but also the understanding the molecular mechanisms of heart diseases.

scholarly journals Investigating LMNA-Related Dilated Cardiomyopathy Using Human Induced Pluripotent Stem Cell-Derived Cardiomyocytes

2021 ◽  
Vol 22 (15) ◽  
pp. 7874
Author(s):  
Yuval Shemer ◽  
Lucy N. Mekies ◽  
Ronen Ben Jehuda ◽  
Polina Baskin ◽  
Rita Shulman ◽  
...  
Keyword(s):  
Stem Cell ◽  
Action Potential ◽  
Dilated Cardiomyopathy ◽  
Pluripotent Stem Cell ◽  
Systolic Function ◽  
Induced Pluripotent Stem Cell ◽  
Left Ventricular ◽  
Lmna Gene ◽  
Beat Rate ◽  
Induced Pluripotent

LMNA-related dilated cardiomyopathy is an inherited heart disease caused by mutations in the LMNA gene encoding for lamin A/C. The disease is characterized by left ventricular enlargement and impaired systolic function associated with conduction defects and ventricular arrhythmias. We hypothesized that LMNA-mutated patients’ induced Pluripotent Stem Cell-derived cardiomyocytes (iPSC-CMs) display electrophysiological abnormalities, thus constituting a suitable tool for deciphering the arrhythmogenic mechanisms of the disease, and possibly for developing novel therapeutic modalities. iPSC-CMs were generated from two related patients (father and son) carrying the same E342K mutation in the LMNA gene. Compared to control iPSC-CMs, LMNA-mutated iPSC-CMs exhibited the following electrophysiological abnormalities: (1) decreased spontaneous action potential beat rate and decreased pacemaker current (If) density; (2) prolonged action potential duration and increased L-type Ca2+ current (ICa,L) density; (3) delayed afterdepolarizations (DADs), arrhythmias and increased beat rate variability; (4) DADs, arrhythmias and cessation of spontaneous firing in response to β-adrenergic stimulation and rapid pacing. Additionally, compared to healthy control, LMNA-mutated iPSC-CMs displayed nuclear morphological irregularities and gene expression alterations. Notably, KB-R7943, a selective inhibitor of the reverse-mode of the Na+/Ca2+ exchanger, blocked the DADs in LMNA-mutated iPSC-CMs. Our findings demonstrate cellular electrophysiological mechanisms underlying the arrhythmias in LMNA-related dilated cardiomyopathy.

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