Human heart disease: lessons from human pluripotent stem cell-derived cardiomyocytes

Giacomelli, Elisa; Mummery, Christine L.; Bellin, Milena

doi:10.1007/s00018-017-2546-5

Cited by 55 publications

(49 citation statements)

References 254 publications

(290 reference statements)

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“…1b). Interestingly, optimal cardiogenic mesoderm induction for all ESC and iPSC lines tested occurred in hHOs that were exposed to lower CHIR99021 concentrations relative to previously reported cardiomyocyte monolayer differentiation protocols, which typically range from 10 to 12 µM CHIR 3,4,6,7,34,39,40 . A 4 µM CHIR99021 exposure resulted in the highest cardiomyocyte content with 64±5% TNNT2 + cells at day 15, versus 9.6±5% and 2.4±2% for 6.6 µM and 8 µM CHIR99021, respectively ( Fig.…”

Section: Differentiation Of Hpscs Into 3d Human Heart Organoids (Hhosmentioning

confidence: 91%

“…hPSCs enable us to recapitulate some of most important developmental steps in vitro to produce specific cardiac cell types with relative ease, high purity and in large amounts 5 . Most of these protocols rely on the controlled stepwise manipulation of the Wnt and BMP signaling pathways using chemical inhibitors 3,6,7 . However, these homogenous cell models are still very far away from the structural and cellular complexity of the tissues and organs they represent (e.g.…”

Section: Introductionmentioning

confidence: 99%

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Generation of Heart Organoids Modeling Early Human Cardiac Development Under Defined Conditions

Ball

Wasserman

et al. 2020

Preprint

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Cardiovascular-related disorders are a significant worldwide health problem. Cardiovascular disease (CVD) is the leading cause of death in developed countries, making up a third of the mortality rate in the US 1 . Congenital heart defects (CHD) affect ~1% of all live births 2 , making it the most common birth defect in humans. Current technologies provide some insight into how these disorders originate but are limited in their ability to provide a complete overview of disease pathogenesis and progression due to their lack of physiological complexity. There is a pressing need to develop more faithful organ-like platforms recapitulating complex in vivo phenotypes to study human development and disease in vitro. Here, we report the most faithful in vitro organoid model of human cardiovascular development to date using human pluripotent stem cells (hPSCs). Our protocol is highly efficient, scalable, shows high reproducibility and is compatible with high-throughput approaches. Furthermore, our hPSC-based heart organoids (hHOs) showed very high similarity to human fetal hearts, both morphologically and in cell-type complexity. hHOs were differentiated using a two-step manipulation of Wnt signaling using chemical inhibitors and growth factors in completely defined media and culture conditions. Organoids were successfully derived from multiple independent hPSCs lines with very similar efficiency. hHOs started beating at ~6 days, were mostly spherical and grew up to ~1 mm in diameter by day 15 of differentiation. hHOs developed sophisticated, interconnected internal chambers and confocal analysis for cardiac markers revealed the presence of all major cardiac lineages, including cardiomyocytes (TNNT2 + ), epicardial cells (WT1 + , TJP + ), cardiac fibroblasts (THY1 + , VIM + ), endothelial cells (PECAM1 + ), and endocardial cells (NFATC1 + ). Morphologically, hHOs developed well-defined epicardial and adjacent myocardial regions and presented a distinct vascular plexus as well as endocardial-lined microchambers. RNA-seq time-course analysis of hHOs, monolayer differentiated iPSCs and fetal human hearts revealed that hHOs recapitulate human fetal heart tissue development better than previously described differentiation protocols 3,4 . hHOs allow higher-order interaction of distinct heart tissues for the first time and display biologically relevant physical and topographical 3D cues that closely resemble the human fetal heart. Our model constitutes a powerful novel tool for discovery and translational studies in human cardiac development and disease.

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Section: Differentiation Of Hpscs Into 3d Human Heart Organoids (Hhosmentioning

confidence: 91%

Section: Introductionmentioning

confidence: 99%

Generation of Heart Organoids Modeling Early Human Cardiac Development Under Defined Conditions

Ball

Wasserman

et al. 2020

Preprint

View full text Add to dashboard Cite

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“…The list of disease models used in the analysis is provided in Supplemental Table S1. This list was generated by initially examining referenced articles from multiple comprehensive reviews published in the field, [9][10][11][12][13][14] . The resulting publication list was then screened for additional relevant publications that were then examined manually.…”

Section: Improving Hipsc Cvd Models Through Genetic Engineeringmentioning

confidence: 99%

Inherited cardiac diseases, pluripotent stem cells, and genome editing combined—the past, present, and future

et al. 2019

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Research on mechanisms underlying monogenic cardiac diseases such as primary arrhythmias and cardiomyopathies has until recently been hampered by inherent limitations of heterologous cell systems, where mutant genes are expressed in noncardiac cells, and physiological differences between humans and experimental animals. Human-induced pluripotent stem cells (hiPSCs) have proven to be a game changer by providing new opportunities for studying the disease in the specific cell type affected, namely the cardiomyocyte. hiPSCs are particularly valuable because not only can they be differentiated into unlimited numbers of these cells, but they also genetically match the individual from whom they were derived. The decade following their discovery showed the potential of hiPSCs for advancing our understanding of cardiovascular diseases, with key pathophysiological features of the patient being reflected in their corresponding hiPSC-derived cardiomyocytes (the past). Now, recent advances in genome editing for repairing or introducing genetic mutations efficiently have enabled the disease etiology and pathogenesis of a particular genotype to be investigated (the present). Finally, we are beginning to witness the promise of hiPSC in personalized therapies for individual patients, as well as their application in identifying genetic variants responsible for or modifying the disease phenotype (the future). In this review, we discuss how hiPSCs could contribute to improving the diagnosis, prognosis, and treatment of an individual with a suspected genetic cardiac disease, thereby developing better risk stratification and clinical management strategies for these potentially lethal but treatable disorders.

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“…To date there have been no reports comparing the in vitro phenotype and physiological properties of cryopreserved and non-frozen hPSC-CMs following replating. Such studies are warranted due to the increasing use of hPSC-CMs for investigating disease mechanisms and in safety pharmacology assays (Brandão et al, 2017;Denning et al, 2016;Giacomelli et al, 2017). For this reason, we have evaluated freshly-derived and cryopreserved cardiomyocytes generated from two different human induced pluripotent stem cell lines (hiPSC-CMs).…”

Section: Introductionmentioning

confidence: 99%

Cryopreservation of human pluripotent stem cell-derived cardiomyocytes is not detrimental to their molecular and functional properties

Brink

Brandão

Grandela

et al. 2019

Preprint

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Human induced pluripotent stem cell-derived cardiomyocytes (hiPSC-CMs) have emerged as a powerful platform for in vitro modelling of cardiac diseases, safety pharmacology, and drug screening.All these applications require large quantities of well-characterised and standardised batches of hiPSC-CMs. Cryopreservation of hiPSC-CMs without affecting their biochemical or biophysical phenotype is essential for facilitating this, but ideally requires the cells being unchanged by the freeze-thaw procedure. We therefore compared the in vitro functional and molecular characteristics of fresh and cryopreserved hiPSC-CMs generated from two independent hiPSC lines. While the frozen hiPSC-CMs exhibited poorer replating than their freshly-derived counterparts, there was no difference in the proportion of cardiomyocytes retrieved from the mixed population when this was factored in.Interestingly, cryopreserved hiPSC-CMs from one line exhibited longer action potential durations.These results provide evidence that cryopreservation does not compromise the in vitro molecular, physiological and mechanical properties of hiPSC-CMs, though can lead to an enrichment in ventricular myocytes. It also validates this procedure for storing hiPSC-CMs, thereby allowing the same batch of hiPSC-CMs to be used for multiple applications and evaluations. 3 Keywords • Human pluripotent stem cell-derived cardiomyocytes • Cryopreservation • Cardiac electrophysiology • Ventricular cardiomyocytes

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Human heart disease: lessons from human pluripotent stem cell-derived cardiomyocytes

Cited by 55 publications

References 254 publications

Generation of Heart Organoids Modeling Early Human Cardiac Development Under Defined Conditions

Generation of Heart Organoids Modeling Early Human Cardiac Development Under Defined Conditions

Inherited cardiac diseases, pluripotent stem cells, and genome editing combined—the past, present, and future

Cryopreservation of human pluripotent stem cell-derived cardiomyocytes is not detrimental to their molecular and functional properties

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