Abstract 30: In vitro Fabrication of Human Cardiac Tissues With Porcine Perfusable Blood Vessels

2016 ◽  
Vol 119 (suppl_1) ◽  
Author(s):  
Akitoshi Inui ◽  
Hidekazu Sekine ◽  
Kazunori Sano ◽  
Izumi Dobashi ◽  
Azumi Yoshida ◽  
...  
Keyword(s):  
Blood Vessels ◽  
Cardiac Tissue ◽  
Severe Heart Failure ◽  
Perfusion Culture ◽  
Cardiac Cell ◽  
Cell Sheets ◽  
Cardiac Tissues ◽  
Bioreactor System ◽  
Human Cardiac Tissue

The definitive treatment of severe heart failure is heart transplantation; however the number of heart transplantation procedures performed in Japan per year ranges from 30-40 due to donor shortage. Therefore, recently other treatments such as ventricular assist device or regenerative therapy by human cardiac tissue engineering have been developed and are considered as appropriate alternatives. We have developed an original technology, which was named cell-sheet based tissue engineering to fabricate functional three-dimensional tissue by layering cell sheets. The utilization of this technique allowed us to successfully engineer thick rat cardiac tissue with perfusable blood vessels in vitro. Here, we demonstrate a technique to engineer human cardiac tissue with perfusable blood vessels using cardiac cell sheets derived from human induced pluripotent stem cells, and porcine small intestine as a vascular bed for perfusion culture. The small intestine was harvested from with a branch of the superior mesenteric artery and vein and underwent mucosal resection after harvested tissue was cut open. To engineer cardiac tissue with perfusable blood vessels, cardiac cell sheets co-cultured with endothelial cells, were triple-layered and then was overlaid on the vascular bed in the bioreactor system. One day after perfusion culture, overlaid cardiac tissues pulsated spontaneously and were synchronized. The cardiac tissue construct was viable tissue without any observable necrosis. Furthermore we examined the possibility of transplantation of the in vitro engineered human cardiac tissue with the connectable host artery and vein. Engineered cardiac tissue was removed from the bioreactor system after 4-day perfusion, and transplanted to another pig heart. The branch of the superior mesenteric artery and vein of the graft were then reconnected to the host internal thoracic artery and vein. When the cardiac tissue reperfused, it began to beat spontaneously after a few minutes. We believe that this method is useful to fabricate functional cardiac tissue and may become an appropriate treatment for severe heart failure.

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 Three-Dimensional Human Cardiac Tissue Engineered by Centrifugation of Stacked Cell Sheets and Cross-Sectional Observation of Its Synchronous Beatings by Optical Coherence Tomography

2017 ◽  
Vol 2017 ◽  
pp. 1-8 ◽  
Author(s):  
Yuji Haraguchi ◽  
Akiyuki Hasegawa ◽  
Katsuhisa Matsuura ◽  
Mari Kobayashi ◽  
Shin-ichi Iwana ◽  
...  
Keyword(s):  
Tissue Engineering ◽  
Optical Coherence Tomography ◽  
Regenerative Medicine ◽  
Cardiac Tissue ◽  
Three Dimensional ◽  
Cardiac Cell ◽  
Fabrication Technique ◽  
Cell Sheets ◽  
Cross Sectional ◽  
Cardiac Tissues

Three-dimensional (3D) tissues are engineered by stacking cell sheets, and these tissues have been applied in clinical regenerative therapies. The optimal fabrication technique of 3D human tissues and the real-time observation system for these tissues are important in tissue engineering, regenerative medicine, cardiac physiology, and the safety testing of candidate chemicals. In this study, for aiming the clinical application, 3D human cardiac tissues were rapidly fabricated by human induced pluripotent stem (iPS) cell-derived cardiac cell sheets with centrifugation, and the structures and beatings in the cardiac tissues were observed cross-sectionally and noninvasively by two optical coherence tomography (OCT) systems. The fabrication time was reduced to approximately one-quarter by centrifugation. The cross-sectional observation showed that multilayered cardiac cell sheets adhered tightly just after centrifugation. Additionally, the cross-sectional transmissions of beatings within multilayered human cardiac tissues were clearly detected by OCT. The observation showed the synchronous beatings of the thicker 3D human cardiac tissues, which were fabricated rapidly by cell sheet technology and centrifugation. The rapid tissue-fabrication technique and OCT technology will show a powerful potential in cardiac tissue engineering, regenerative medicine, and drug discovery research.

Biophysical stimulation for in vitro engineering of functional cardiac tissues

2017 ◽  
Vol 131 (13) ◽  
pp. 1393-1404 ◽  
Author(s):  
Anastasia Korolj ◽  
Erika Yan Wang ◽  
Robert A. Civitarese ◽  
Milica Radisic
Keyword(s):  
Cardiac Tissue ◽  
Culture Media ◽  
Culture Conditions ◽  
Lineage Specification ◽  
Cardiac Tissues ◽  
Cardiac Lineage ◽  
And Function ◽  
Tissue Models

Engineering functional cardiac tissues remains an ongoing significant challenge due to the complexity of the native environment. However, our growing understanding of key parameters of the in vivo cardiac microenvironment and our ability to replicate those parameters in vitro are resulting in the development of increasingly sophisticated models of engineered cardiac tissues (ECT). This review examines some of the most relevant parameters that may be applied in culture leading to higher fidelity cardiac tissue models. These include the biochemical composition of culture media and cardiac lineage specification, co-culture conditions, electrical and mechanical stimulation, and the application of hydrogels, various biomaterials, and scaffolds. The review will also summarize some of the recent functional human tissue models that have been developed for in vivo and in vitro applications. Ultimately, the creation of sophisticated ECT that replicate native structure and function will be instrumental in advancing cell-based therapeutics and in providing advanced models for drug discovery and testing.

In Vitro Studies of Human Cardiac Tissue

1970 ◽  
Vol 33 (5) ◽  
pp. 480-480
Author(s):  
RICHARD A. JENSEN
Keyword(s):  
Cardiac Tissue ◽  
In Vitro Studies ◽  
Human Cardiac Tissue

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 A three-dimensional hybrid pacemaker electrode seamlessly integrates into engineered, functional human cardiac tissue in vitro

2018 ◽  
Vol 8 (1) ◽  
Author(s):  
Tobias Weigel ◽  
Tobias Schmitz ◽  
Tobias Pfister ◽  
Sabine Gaetzner ◽  
Maren Jannasch ◽  
...  
Keyword(s):  
Cardiac Tissue ◽  
Three Dimensional ◽  
Human Cardiac Tissue

scholarly journals Chronic optical pacing conditioning of h-iPSC engineered cardiac tissues

2019 ◽  
Vol 10 ◽  
pp. 204173141984174 ◽  
Author(s):  
Marc Dwenger ◽  
William J Kowalski ◽  
Fei Ye ◽  
Fangping Yuan ◽  
Joseph P Tinney ◽  
...  
Keyword(s):  
Stem Cell ◽  
Electrical Stimulation ◽  
Pluripotent Stem Cell ◽  
Cardiac Tissue ◽  
Induced Pluripotent Stem Cell ◽  
Mechanical Stretch ◽  
In Vitro Models ◽  
Cardiac Tissues ◽  
Induced Pluripotent

The immaturity of human induced pluripotent stem cell derived engineered cardiac tissues limits their ability to regenerate damaged myocardium and to serve as robust in vitro models for human disease and drug toxicity studies. Several chronic biomimetic conditioning protocols, including mechanical stretch, perfusion, and/or electrical stimulation promote engineered cardiac tissue maturation but have significant technical limitations. Non-contacting chronic optical stimulation using heterologously expressed channelrhodopsin light-gated ion channels, termed optogenetics, may be an advantageous alternative to chronic invasive electrical stimulation for engineered cardiac tissue conditioning. We designed proof-of-principle experiments to successfully transfect human induced pluripotent stem cell derived engineered cardiac tissues with a desensitization resistant, chimeric channelrhodopsin protein, and then optically paced engineered cardiac tissues to accelerate maturation. We transfected human induced pluripotent stem cell engineered cardiac tissues using an adeno-associated virus packaged chimeric channelrhodopsin and then verified optically paced by whole cell patch clamp. Engineered cardiac tissues were then chronically optically paced above their intrinsic beat rates in vitro from day 7 to 14. Chronically optically paced resulted in improved engineered cardiac tissue electrophysiological properties and subtle changes in the expression of some cardiac relevant genes, though active force generation and histology were unchanged. These results validate the feasibility of a novel chronically optically paced paradigm to explore non-invasive and scalable optically paced–induced engineered cardiac tissue maturation strategies.

scholarly journals SARS-CoV-2 infects and induces cytotoxic effects in human cardiomyocytes

2020 ◽  
Author(s):  
Denisa Bojkova ◽  
Julian Wagner ◽  
Mariana Shumliakivska ◽  
Galip Aslan ◽  
Umber Saleem ◽  
...  
Keyword(s):  
Cardiac Tissue ◽  
Cardiac Injury ◽  
Transcriptional Response ◽  
Human Cardiomyocytes ◽  
Global Pandemic ◽  
Virus Rna ◽  
Human Cardiac Tissue ◽  
Induced Pluripotent ◽  
Apoptotic Effects

Background: The coronavirus disease 2019 (COVID-19) is caused by severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2) and has emerged as global pandemic. SARS-CoV-2 infection can lead to elevated markers of cardiac injury associated with higher risk of mortality in COVID-19 patients. It is unclear whether cardiac injury may have been caused by direct infection of cardiomyocytes or is mainly secondary to lung injury and inflammation. Here we investigate whether human cardiomyocytes are permissive for SARS-CoV-2 infection. Methods: Infection was induced by two strains of SARS-CoV-2 (FFM1 and FFM2) in human induced pluripotent stem cells-derived cardiomyocytes (hiPS-CM) and in two models of human cardiac tissue. Results: We show that SARS-CoV-2 infects hiPS-CM as demonstrated by detection of intracellular double strand viral RNA and viral spike glycoprotein protein expression. Increasing concentrations of virus RNA are detected in supernatants of infected cardiomyocytes, which induced infections in CaCo-2 cell lines documenting productive infections. SARS-COV-2 infection induced cytotoxic and pro-apoptotic effects and abolished cardiomyocyte beating. RNA sequencing confirmed a transcriptional response to viral infection as demonstrated by the up-regulation of genes associated with pathways related to viral response and interferon signaling, apoptosis and reactive oxygen stress. SARS-CoV-2 infection and cardiotoxicity was confirmed in a iPS-derived human 3D cardiosphere tissue models. Importantly, viral spike protein and viral particles were detected in living human heart slices after infection with SARS-CoV-2. Conclusions: The demonstration that cardiomyocytes are permissive for SARS-CoV-2 infection in vitro warrants the further in depth monitoring of cardiotoxic effects in COVID-19 patients.

High-aspect-ratio water-dispersed gold nanowires incorporated within gelatin methacrylate hydrogels for constructing cardiac tissues in vitro

2020 ◽  
Vol 8 (32) ◽  
pp. 7213-7224 ◽  
Author(s):  
Xiao-Pei Li ◽  
Kai-Yun Qu ◽  
Feng Zhang ◽  
Han-Ning Jiang ◽  
Ning Zhang ◽  
...  
Keyword(s):  
Aspect Ratio ◽  
High Aspect Ratio ◽  
Cardiac Tissue ◽  
Gold Nanowires ◽  
Tissue Formation ◽  
Cardiac Tissues ◽  
Cardiomyocyte Culture ◽  
Dispersed Gold

The prepared high-aspect-ratio water-dispersed gold nanowires are incorporated into GeIMA hydrogels for cardiomyocyte culture and micro-cardiac tissue formation.

scholarly journals Cardiovascular tissue regeneration system based on multiscale scaffolds comprising double-layered hydrogels and fibers

2020 ◽  
Vol 10 (1) ◽  
Author(s):  
Yun-Min Kook ◽  
Soonjae Hwang ◽  
Hyerim Kim ◽  
Ki-Jong Rhee ◽  
Kangwon Lee ◽  
...  
Keyword(s):  
Stem Cells ◽  
Blood Vessels ◽  
Cardiac Tissue ◽  
Umbilical Vein ◽  
Peripheral Region ◽  
Regeneration System ◽  
Alginate Hydrogel ◽  
Cardiovascular Tissue ◽  
Encapsulated Cells

AbstractWe report a technique to reconstruct cardiovascular tissue using multiscale scaffolds incorporating polycaprolactone fibers with double-layered hydrogels comprising fibrin hydrogel surrounded by secondary alginate hydrogel. The scaffolds compartmentalized cells into the core region of cardiac tissue and the peripheral region of blood vessels to construct cardiovascular tissue, which was accomplished by a triple culture system of adipose-derived mesenchymal stem cells (ADSCs) with C2C12 myoblasts on polycaprolactone (PCL) fibers along with human umbilical vein endothelial cells (HUVECs) in fibrin hydrogel. The secondary alginate hydrogel prevented encapsulated cells from migrating outside scaffold and maintained the scaffold structure without distortion after subcutaneous implantation. According to in vitro studies, resultant scaffolds promoted new blood vessel formation as well as cardiomyogenic phenotype expression of ADSCs. Cardiac muscle-specific genes were expressed from stem cells and peripheral blood vessels from HUVECs were also successfully developed in subcutaneously implanted cell-laden multiscale scaffolds. Furthermore, the encapsulated stem cells modulated the immune response of scaffolds by secreting anti-inflammatory cytokines for successful tissue construction. Our study reveals that multiscale scaffolds can be promising for the remodeling and transplantation of cardiovascular tissue.

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