scholarly journals Human iPSC-Derived Cardiomyocytes are Susceptible to SARS-CoV-2 Infection

2020 ◽  
Author(s):  
Arun Sharma ◽  
Gustavo Garcia ◽  
Vaithilingaraja Arumugaswami ◽  
Clive N. Svendsen
Keyword(s):  
Inflammatory Responses ◽  
Antiviral Drug ◽  
Viral Myocarditis ◽  
Induced Pluripotent Stem Cell ◽  
Cardiac Complications ◽  
Human Cardiomyocytes ◽  
Screening Platform ◽  
Induced Pluripotent ◽  
Human Ipsc

SUMMARYCoronavirus disease 2019 (COVID-19) is a viral pandemic caused by the severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2). COVID-19 is predominantly defined by respiratory symptoms, but cardiac complications including arrhythmias, heart failure, and viral myocarditis are also prevalent. Although the systemic ischemic and inflammatory responses caused by COVID-19 can detrimentally affect cardiac function, the direct impact of SARS-CoV-2 infection on human cardiomyocytes is not well-understood. We used human induced pluripotent stem cell-derived cardiomyocytes (hiPSC-CMs) as a model system to examine the mechanisms of cardiomyocyte-specific infection by SARS-CoV-2. Microscopy and immunofluorescence demonstrated that SARS-CoV-2 can enter and replicate within hiPSC-CMs, localizing at perinuclear locations within the cytoplasm. Viral cytopathic effect induced hiPSC-CM apoptosis and cessation of beating after 72 hours of infection. These studies show that SARS-CoV-2 can infect hiPSC-CMs in vitro, establishing a model for elucidating the mechanisms of infection and potentially a cardiac-specific antiviral drug screening platform.

scholarly journals Maturation of human induced pluripotent stem cell-derived cardiomyocytes for modeling hypertrophic cardiomyopathy

2020 ◽  
Author(s):  
Walter E. Knight ◽  
Yingqiong Cao ◽  
Ying-Hsi Lin ◽  
Genevieve C. Sparagna ◽  
Betty Bai ◽  
...  
Keyword(s):  
Cardiovascular Disease ◽  
Stem Cell ◽  
Pluripotent Stem Cell ◽  
Induced Pluripotent Stem Cell ◽  
Simple Method ◽  
Human Cardiomyocytes ◽  
Pathological Hypertrophy ◽  
Induced Pluripotent ◽  
Metabolic Behavior

AbstractRationaleHuman induced pluripotent stem cell derived cardiomyocytes (hiPSC-CMs) are a powerful platform for biomedical research. However, they are immature, which is a barrier to modeling adult-onset cardiovascular disease.ObjectiveWe sought to develop a simple method which could drive cultured hiPSC-CMs towards maturity across a number of phenotypes.Methods and resultsCells were cultured in fatty acid-based media and plated on micropatterned surfaces to promote alignment and elongation. These cells display many characteristics of adult human cardiomyocytes, including elongated cell morphology, enhanced maturity of sarcomeric structures, metabolic behavior, and increased myofibril contractile force. Most notably, hiPSC-CMs cultured under optimal maturity-inducing conditions recapitulate the pathological hypertrophy caused by either a pro-hypertrophic agent or genetic mutations.ConclusionsThe more mature hiPSC-CMs produced by the methods described here will serve as a useful in vitro platform for characterizing cardiovascular disease.

scholarly journals Tension-driven multi-scale self-organisation in human iPSC-derived muscle fibers

2021 ◽  
Author(s):  
Qiyan Mao ◽  
Achyuth Acharya ◽  
Alejandra Rodriguez-delaRosa ◽  
Fabio Marchiano ◽  
Benoit Dehapiot ◽  
...  
Keyword(s):  
Induced Pluripotent Stem Cell ◽  
Human Muscle ◽  
Mechanical Tension ◽  
In Vitro System ◽  
Multi Scale ◽  
Self Organisation ◽  
Induced Pluripotent ◽  
Sarcomere Assembly ◽  
Human Ipsc

Human muscle is a hierarchically organised tissue with its contractile cells called myofibers packed into large myofiber bundles. Each myofiber contains periodic myofibrils built by hundreds of contractile sarcomeres that generate large mechanical forces. To better understand the mechanisms that coordinate human muscle morphogenesis from tissue to molecular scales, we adopted a simple in vitro system using induced pluripotent stem cell-derived human myogenic precursors. When grown on an unrestricted two-dimensional substrate, developing myofibers spontaneously align and self-organise into higher-order myofiber bundles, which grow and consolidate to stable sizes. Following a transcriptional boost of sarcomeric components, myofibrils assemble into chains of periodic sarcomeres that emerge across the entire myofiber. By directly probing tension we found that tension build-up precedes sarcomere assembly and increases within each assembling myofibril. Furthermore, we found that myofiber ends stably attach to other myofibers using integrin-based attachments and thus myofiber bundling coincides with stable myofiber bundle attachment in vitro. A failure in stable myofiber attachment results in a collapse followed by a disassembly of the myofibrils. Overall, our results strongly suggest that mechanical tension across sarcomeric components as well as between differentiating myofibers is key to coordinate the multi-scale self-organisation of muscle morphogenesis.

scholarly journals Human Erbb2-induced Erk Activity Robustly Stimulates Cycling and Functional Remodeling of Rat and Human Cardiomyocytes

2020 ◽  
Author(s):  
Nicholas Strash ◽  
Sophia DeLuca ◽  
Geovanni Janer Carattini ◽  
Soon Chul Heo ◽  
Ryne Gorsuch ◽  
...  
Keyword(s):  
Ventricular Myocytes ◽  
Induced Pluripotent Stem Cell ◽  
Neonatal Rat ◽  
Mutant Form ◽  
Cardiac Tissues ◽  
Human Cardiomyocytes ◽  
Heart Repair ◽  
Induced Pluripotent ◽  
Species Specific

Multiple mitogenic pathways capable of promoting mammalian cardiomyocyte (CM) proliferation have been identified as potential candidates for functional heart repair following myocardial infarction. However, it is unclear whether the effects of these mitogens are species-specific and how they directly compare in the same cardiac setting. Here, we examined how CM-specific lentiviral expression of various candidate mitogens affects human induced pluripotent stem cell-derived cardiomyocytes (hiPSC-CMs) and neonatal rat ventricular myocytes (NRVMs) in vitro. In 2D-cultured CMs from both species, and in highly mature 3D-engineered cardiac tissues generated from NRVMs, a constitutively-active mutant form of the human gene Erbb2 (cahErbb2) was the most potent tested mitogen. Persistent expression of cahErbb2 induced CM proliferation, sarcomere loss, and remodeling of tissue structure and function, which were attenuated by small molecule inhibitors of Erk signaling. These results suggest transient activation of Erbb2/Erk axis in cardiomyocytes as a potential strategy for regenerative heart repair.

scholarly journals Topologically controlled circuits of human iPSC-derived neurons for electrophysiology recordings

2021 ◽  
Author(s):  
Sophie Girardin ◽  
Blandine Clément ◽  
Stephan J. Ihle ◽  
Sean Weaver ◽  
Jana B. Petr ◽  
...  
Keyword(s):  
Information Processing ◽  
Neurological Disorders ◽  
Induced Pluripotent Stem Cell ◽  
Animal Origin ◽  
Great Promise ◽  
Human Neurons ◽  
Induced Pluripotent ◽  
Human Ipsc ◽  
The Brain

Bottom-up neuroscience, which consists of building and studying controlled networks of neurons in vitro, is a promising method to investigate information processing at the neuronal level. However, in vitro studies tend to use cells of animal origin rather than human neurons, leading to conclusions that might not be generalizable to humans and limiting the possibilities for relevant studies on neurological disorders. Here we present a method to build arrays of topologically controlled circuits of human induced pluripotent stem cell (iPSC)-derived neurons. The circuits consist of 4 to 50 neurons with mostly unidirectional connections, confined by microfabricated polydimethylsiloxane (PDMS) membranes. Such circuits were characterized using optical imaging and microelectrode arrays (MEAs). Electrophysiology recordings were performed on circuits of human iPSC-derived neurons for at least 4.5 months. We believe that the capacity to build small and controlled circuits of human iPSC-derived neurons holds great promise to better understand the fundamental principles of information processing and storing in the brain.

scholarly journals Honing the Double-Edged Sword: Improving Human iPSC-Microglia Models

2020 ◽  
Vol 11 ◽  
Author(s):  
Anne Hedegaard ◽  
Szymon Stodolak ◽  
William S. James ◽  
Sally A. Cowley
Keyword(s):  
Extracellular Matrix ◽  
Stem Cell ◽  
Pluripotent Stem Cell ◽  
Induced Pluripotent Stem Cell ◽  
Media Composition ◽  
Physiological Relevance ◽  
Key Features ◽  
Induced Pluripotent ◽  
Human Ipsc

Human induced Pluripotent Stem Cell (hiPSC) models are a valuable new tool for research into neurodegenerative diseases. Neuroinflammation is now recognized as a key process in neurodegenerative disease and aging, and microglia are central players in this. A plethora of hiPSC-derived microglial models have been published recently to explore neuroinflammation, ranging from monoculture through to xenotransplantation. However, combining physiological relevance, reproducibility, and scalability into one model is still a challenge. We examine key features of the in vitro microglial environment, especially media composition, extracellular matrix, and co-culture, to identify areas for improvement in current hiPSC-microglia models.

scholarly journals Human Erbb2-induced Erk activity robustly stimulates cycling and functional remodeling of rat and human cardiomyocytes

eLife ◽  
2021 ◽  
Vol 10 ◽  
Author(s):  
Nicholas Strash ◽  
Sophia DeLuca ◽  
Geovanni L Janer Carattini ◽  
Soon Chul Heo ◽  
Ryne Gorsuch ◽  
...  
Keyword(s):  
Ventricular Myocytes ◽  
Induced Pluripotent Stem Cell ◽  
Neonatal Rat ◽  
Mutant Form ◽  
Cardiac Tissues ◽  
Human Cardiomyocytes ◽  
Heart Repair ◽  
Induced Pluripotent ◽  
Species Specific

Multiple mitogenic pathways capable of promoting mammalian cardiomyocyte (CM) proliferation have been identified as potential candidates for functional heart repair following myocardial infarction. However, it is unclear whether the effects of these mitogens are species-specific and how they directly compare in the same cardiac setting. Here, we examined how CM-specific lentiviral expression of various candidate mitogens affects human induced pluripotent stem cell-derived CMs (hiPSC-CMs) and neonatal rat ventricular myocytes (NRVMs) in vitro. In 2D-cultured CMs from both species, and in highly mature 3D-engineered cardiac tissues generated from NRVMs, a constitutively-active mutant form of the human gene Erbb2 (cahErbb2) was the most potent tested mitogen. Persistent expression of cahErbb2 induced CM proliferation, sarcomere loss, and remodeling of tissue structure and function, which were attenuated by small molecule inhibitors of Erk signaling. These results suggest transient activation of Erbb2/Erk axis in cardiomyocytes as a potential strategy for regenerative heart repair.

scholarly journals CD49f is a novel marker to purify functional human iPSC-derived astrocytes

2019 ◽  
Author(s):  
Lilianne Barbar ◽  
Tanya Jain ◽  
Matthew Zimmer ◽  
Ilya Kruglikov ◽  
Suzanne R. Burstein ◽  
...  
Keyword(s):  
Neurodegenerative Diseases ◽  
Glutamate Uptake ◽  
Induced Pluripotent Stem Cell ◽  
Characteristic Functions ◽  
Patient Specific ◽  
The Central Nervous System ◽  
Induced Pluripotent ◽  
Complex Culture ◽  
Human Ipsc

ABSTRACTAstrocytes play a central role in the central nervous system (CNS), maintaining brain homeostasis, providing metabolic support to neurons, regulating connectivity of neural circuits, and controlling blood flow as an integral part of the blood-brain barrier. They have been increasingly implicated in the mechanisms of neurodegenerative diseases, prompting a greater need for methods that enable their study. The advent of human induced pluripotent stem cell (iPSC) technology has made it possible to generate patient-specific astrocytes and CNS cells using protocols developed by our team and others as valuable disease models. Yet isolating astrocytes from primary specimens or from in vitro mixed cultures for downstream analyses has remained challenging. To address this need, we performed a screen for surface markers that allow FACS sorting of astrocytes. Here we demonstrate that CD49f is an effective marker for sorting functional human astrocytes. We sorted CD49f+ cells from a protocol we previously developed that generates a complex culture of oligodendrocytes, neurons and astrocytes from iPSCs. CD49f+-purified cells express all canonical astrocyte markers and perform characteristic functions, such as neuronal support and glutamate uptake. Of particular relevance to neurodegenerative diseases, CD49f+ astrocytes can be stimulated to take on an A1 neurotoxic phenotype, in which they secrete pro-inflammatory cytokines and show an impaired ability to support neuronal maturation. This study establishes a novel marker for isolating functional astrocytes from complex CNS cell populations, strengthening the use of iPSC-astrocytes for the study of their regulation and dysregulation in neurodegenerative diseases.

scholarly journals Differentiation, Evaluation, and Application of Human Induced Pluripotent Stem Cell–Derived Endothelial Cells

2017 ◽  
Vol 37 (11) ◽  
pp. 2014-2025 ◽  
Author(s):  
Yang Lin ◽  
Chang-Hyun Gil ◽  
Mervin C. Yoder
Keyword(s):  
Endothelial Cells ◽  
Stem Cell ◽  
Pluripotent Stem Cell ◽  
Disease Modeling ◽  
Induced Pluripotent Stem Cell ◽  
Patient Specific ◽  
Induced Pluripotent ◽  
Human Ipsc

The emergence of induced pluripotent stem cell (iPSC) technology paves the way to generate large numbers of patient-specific endothelial cells (ECs) that can be potentially delivered for regenerative medicine in patients with cardiovascular disease. In the last decade, numerous protocols that differentiate EC from iPSC have been developed by many groups. In this review, we will discuss several common strategies that have been optimized for human iPSC-EC differentiation and subsequent studies that have evaluated the potential of human iPSC-EC as a cell therapy or as a tool in disease modeling. In addition, we will emphasize the importance of using in vivo vessel-forming ability and in vitro clonogenic colony–forming potential as a gold standard with which to evaluate the quality of human iPSC-EC derived from various protocols.

Abstract 439: Modeling Diabetic Cardiomyopathy Model Using Human Induced Pluripotent Stem Cell-derived Cardiomyocytes

2016 ◽  
Vol 119 (suppl_1) ◽  
Author(s):  
Yuichi Horikoshi ◽  
Anna Williams ◽  
Xiaowen Bai ◽  
Yasheng Yan ◽  
Wai-Meng Kwok ◽  
...  
Keyword(s):  
Stem Cell ◽  
Diabetic Cardiomyopathy ◽  
Pluripotent Stem Cell ◽  
Induced Pluripotent Stem Cell ◽  
Cardiac Complications ◽  
Diabetic Patients ◽  
Α Subunit ◽  
Maturation Medium ◽  
Induced Pluripotent

Background: Diabetic cardiomyopathy (DCM) is one of the major cardiac complications in diabetic patients. However, the underlying mechanisms and pathogenesis of diabetic cardiomyopathy are only partially understood and the specific therapeutic target remains unknown. Here, we developed an in vitro diabetic cardiomyopathy model through simple metabolic manipulation using human induced pluripotent stem cell (iPSC)-derived cardiomyocytes (iPSC-CMs). Methods: After 30 days from initial differentiation, iPSC-CMs were cultured with maturation medium (containing fatty acid but no glucose) for 3 days to obtain matured cardiomyocyte properties. Maturation extent of iPSC-CMs was analyzed using patch-clamp to analyze electrophysiological properties and RT-PCR to analyze cardiomyocyte-specific structure proteins and ion channel expression. Matured iPSC-CMs were then treated with 11 mM glucose for 2 days. 11 mM glucose represents mild glucose conditions in diabetic patients. Cardiomyopathy phenotype of iPSC-CMs was evaluated by analyzing brain natriuretic peptide (BNP) secretion from iPSC-CMs using ELISA. Result: Differentiated iPSCs-CMs were immature. Following 3 day culture in maturation medium, iPSC-CMs exhibited the cardiomyocyte maturation characteristics: 1) an increase of contractility with diminished automaticity, suggesting progressive maturation toward adult cardiomyocytes, 2) the lower maximal diastolic potential and increased amplitude, indicating functional maturation, and 3) an upregulation of the following gene expression: sarcomeric proteins (e.g., TNNI3, MYL2 and MYL3) and potassium channel α-subunit genes (KCNJ12 and KCNJ4). Matured iPSC-CMs treated with 11 mM glucose showed increase in cell size and BNP secretion (17-fold increase vs. control cells without glucose, p<0.01). These results suggested that cardiomyocytes exposure to the maturation medium followed by diabetic milieu of glucose recapitulate DCM in vitro . Conclusion: We conclude that the phenotype of DCM can be modeled using cultured iPSC-derived cardiomyocytes through diabetic extracellular milieu. This clinically relevant stem cell model would be a powerful platform to study novel mechanisms of DCM and the specific therapeutic targets.

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