Name: implantation of hipsc-derived cardiac-muscle patches after myocardial injury in a guinea pig model
Uploaded: 2019-03-18T13:00:00
Description: Watch this Scientific Journal Video about Implantation of hiPSC-derived Cardiac-muscle Patches after Myocardial Injury in a Guinea Pig Model at JoVE.com

No funding was received for this study

Animals received humane care in compliance with the Guide for the Principles of Laboratory Animals, prepared by the Institute of Laboratory Animal Resources, and published by the National Institutes of Health. All animal protocols were approved by the responsible local authority (&#8216;&#8216;Amt f&#252;r Gesundheit und Verbraucherschutz, Hansestadt Hamburg&#8217;&#8217;/ Animal protocol # 109/16).
1. Obtain Animals
<ol>
	<li>Commercially obtain female Guinea pigs weighing 500&#8211;600 g.<...

<ol>
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A. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Cardiac+applications+for+human+pluripotent+stem+cells.">Cardiac applications for human pluripotent stem cells.</a> Current pharmaceutical design. 15, 2791-2806 (2009).</li><li>Shiba, Y., et al. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Human+ES-cell-derived+cardiomyocytes+electrically+couple+and+suppress+arrhythmias+in+injured+hearts.">Human ES-cell-derived cardiomyocytes electrically couple and suppress arrhythmias in injured hearts.</a> Nature. 489, 322-325 (2012).</li><li>Chong, J. J., et al. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Human+embryonic-stem-cell-derived+cardiomyocytes+regenerate+non-human+primate+hearts.">Human embryonic-stem-cell-derived cardiomyocytes regenerate non-human primate hearts.</a> Nature. 510, 273-277 (2014).</li><li>Kattman, S. J., et al. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Stage-specific+optimization+of+activin/nodal+and+BMP+signaling+promotes+cardiac+differentiation+of+mouse+and+human+pluripotent+stem+cell+lines.">Stage-specific optimization of activin/nodal and BMP signaling promotes cardiac differentiation of mouse and human pluripotent stem cell lines.</a> Cell stem cell. 8, 228-240 (2011).</li><li>Watanabe, T., Rautaharju, P. M., McDonald, T. 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A variety of small animal models are available to study the effect that cell transplantation exerts on injured hearts9,10,11. We chose a guinea pig model because of all small animal models its (electro)physiology resembles most closely that of humans. The advantages of small animal models are simple housing, manageable costs, and few workforces. In comparison with mice and rats, guinea pigs&#180; cardiac (electro)physiology is m...

The number of patients with heart failure is increasing in our aging population. For end-stage heart failure, orthotopic heart transplantation is the only curative treatment option. However, especially in European countries, there is an increasing organ donor shortage. Therefore, alternative treatment options are necessary. Recent achievements in mechanical circulatory support are promising, but especially in the long-term run, limited by its side effects like bleeding, pump thrombosis and infectious complications1.
The endogenous regeneration capacity of the adult human heart is extremely limited. Therefore, cardiac regeneration therapies might become an alternative treatment option for end-stage heart failure patients2,3. Different techniques including stem cell-based cell injection or tissue engineering approaches have been described3,4,5.
Human induced pluripotent stem cells (hiPSC), as well as human embryonic stem cells (hESC) can be effectively differentiated to spontaneously beating human cardiomyocytes6, which has been a major achievement in the field of cardiac regenerative therapies.
To replace myocardium after a myocardial infarction and to improve the function of a failing heart, survival of a suitable number of cardiomyocytes and their mechanical and electrical coupling with the native heart is essential. To investigate the potential of cardiac regenerative therapies with human iPS cell derived cardiomyocytes, a suitable research model is needed. The ideal model should be cost-effective and have a human-like physiology and electrophysiology. Large animal models like pigs would be ideal from that point of view, however, those experiments are very expensive and large amounts of cardiomyocytes would be necessary to replace a relevant number of cardiomyocytes in order to see effects on the left ventricular function in a pig infarction model.
To answer elementary biological questions towards human cell-based cardiac regeneration, e.g., cell survival, vascularization, and electrical coupling, small animal models are more suitable. From the available small animal models, the guinea pig is the most useful species, as compared to rats and mice, as their electrophysiology more closely resembles the situation in humans7. In this guinea pig model, we induced a transmural cryoinjury of the left ventricle. One week after induction of myocardial infarction implantation of a three-dimensional, spontaneously beating hiPS-cell derived cardiomyocyte patch was performed. Cardiomyocyte cell survival was evaluated 28 days after implantation by histological examination.

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implantation of hipsc-derived cardiac-muscle patches after myocardial injury in a guinea pig model

Due to the limited regeneration capacity of the heart in adult mammals, myocardial infarction results in an irreversible loss of cardiomyocytes. This loss of relevant amounts of heart muscle mass can lead to the heart failure. Besides heart transplantation, there is no curative treatment option for the end-stage heart failure. In times of organ donor shortage, organ independent treatment modalities are needed. Left-ventricular assist devices are a promising therapy option, however, especially as destination therapy, limited by its side-effects like stroke, infections and bleedings. In recent years, several cardiac repair strategies including stem cell injection, cardiac progenitors or myocardial tissue engineering have been investigated. Recent improvements in cell biology allow for the differentiation of large amounts of cardiomyocytes derived from human induced pluripotent stem cells (iPSC). One of the cardiac repair strategies currently under evaluation is to transplant artificial heart tissue. Engineered heart tissue (EHT) is a three-dimensional in vitro created cardiomyocyte network, with functional properties of native heart tissue. We have created EHT-patches from hiPSC derived cardiomyocytes. Here we present a protocol for the induction of left ventricular myocardial cryoinjury in a guinea pig, followed by implantation of hiPSC derived EHT on the left ventricular wall.

This guinea pig model is a suitable model to investigate cardiac regeneration after implantation of hiPSC derived EHT-patches. It reproducibly leads to large transmural myocardial injuries. Scar size is evaluated by histology four weeks after cryoinjury. Mason trichrome staining reveals large transmural scars (Figure 2). Scar size was similar over a large number of injured animals reflecting a high degree of reproducibility8. On average 25% of the left ventricular myo...

Watch this Scientific Journal Video about Implantation of hiPSC-derived Cardiac-muscle Patches after Myocardial Injury in a Guinea Pig Model at JoVE.com

Implantation of hiPSC-derived Cardiac-muscle Patches after Myocardial Injury in a Guinea Pig Model

Here we present a protocol for the induction of left ventricular cryoinjury followed by the implantation of a cardiac muscle patch, derived from human iPS-cell cardiomyocytes in a guinea pig model.

Due to the limited regeneration capacity of the heart in adult mammals, myocardial infarction results in an irreversible loss of cardiomyocytes. This loss ...

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