The impact of in vitro cell culture duration on the maturation of human cardiomyocytes derived from induced pluripotent stem cells of myogenic origin

Lewandowski, Jarosław; Rozwadowska, Natalia; Kolanowski, Tomasz; Malcher, Agnieszka; Zimna, Agnieszka; Rugowska, Anna; Fiedorowicz, Katarzyna; Łabędź, Wojciech; Kubaszewski, Łukasz; Chojnacka, Katarzyna; Bednarek-Rajewska, Katarzyna; Majewski, Przemysław; Kurpisz, Maciej

doi:10.1177/0963689718779346

Cited by 63 publications

(60 citation statements)

References 59 publications

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“…This is in line with results from other studies where extended culture of the CMs also showed enhanced maturation of the cells (23)(24)(25). The difference in response related to time in culture can probably be one explanation of why there is a discrepancy among the data reported previously regarding hypertrophic responses in stem cell-derived CMs (26,27).…”

Section: Discussionsupporting

confidence: 90%

Cardiac Hypertrophy in a Dish: A Human Stem Cell Based Model

Johansson¹,

Ulfenborg²,

Andersson³

et al. 2019

Preprint

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Cardiac hypertrophy is an important and independent risk factor for the development of heart failure. To better understand the mechanisms and regulatory pathways involved in cardiac hypertrophy, there is a need for improved in vitro models. In this study, we investigated how hypertrophic stimulation affected human induced pluripotent stem cell (iPSC)-derived cardiomyocytes (CMs). The cells were stimulated with endothelin-1 (ET-1) for 8, 24, 48, 72, or 96h. Parameters including cell size, ANP-, proBNP-, and lactate concentration were analyzed. Moreover, transcriptional profiling using RNA-sequencing was performed to identify differentially expressed genes following ET-1 stimulation. The results show that the CMs increase in size by approximately 13% when exposed to ET-1 in parallel to increases in ANP and proBNP protein and mRNA levels. Furthermore, the lactate concentration in the media was significantly increased indicating that the CMs consume more glucose, a hallmark of cardiac hypertrophy. Using RNA-seq, a hypertrophic gene expression pattern was also observed in the stimulated CMs. Taken together, these results show that hiPSC-derived CMs stimulated with ET-1 display a hypertrophic response. The results from this study also provide new molecular insights about the underlying mechanisms of cardiac hypertrophy and may help accelerate the development of new drugs against this condition. hypertrophy including adverse gene expression profile, increase in ANP and BNP protein levels, and increased lactate production due to a higher glucose consumption (8-10). This type of cardiac hypertrophy is induced by conditions such as chronic hypertension, aortic stenosis, myocardial infarction, or gene mutations. Until recently, only animal-based models have been available to study cardiac hypertrophy in a pre-clinical setting. These models are in many ways useful, but there are significant differences between human-and animal-cardiovascular systems, including stress response and ion channel expression, that makes translation to the human situation challenging (11). Novel technologies, based on human stem cells, could offer clinically relevant in vitro-based alternatives.Human pluripotent stem cells (hPSCs) have a unique capability to self-renew and differentiate into all cell types in the body (12). These features make them useful for various in vitro applications, such as toxicity testing and disease modeling. In particular, hPSC-derived CMs have proven to be useful in many in vitro assays (13)(14)(15). Human cell-based models are anticipated to provide alternatives to the use of animal models for studies of cardiac hypertrophy mechanisms. Besides providing systems that are scalable and may improve the translation of the results to the clinical situation, the availability of human cell-based models can also help to reduce the need for animal experiments.Cardiac hypertrophy can be induced by different methods in vitro, where the most commonly used are neurohormonal stimulation and physical stretching. The neurohormonal appr...

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Section: Discussionsupporting

confidence: 90%

Cardiac Hypertrophy in a Dish: A Human Stem Cell Based Model

Johansson¹,

Ulfenborg²,

Andersson³

et al. 2019

Preprint

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“…The advantages of I K1 -enhanced iPSC-CM include the absence of spontaneous beating (and therefore allow the possibility of controlling electrical pacing), and a resting membrane potential close to physiological values (i.e., −80 mV) [167]. Other strategies to improve the maturation of hiPSC-CMs include long term culture [168] and specific biochemical methods such as using the hormone triiodothyronine [169] or fatty acids [170]. The bioengineering approach, which consists of generating engineered three-dimensional human cardiac tissue, and applying electrical stimulation or mechanical strain to improve both structural and functional maturation, shows great promise [171].…”

Section: Limitations Of Using Human Induced Pluripotent Stem Cell-dermentioning

confidence: 99%

The Emergence of Human Induced Pluripotent Stem Cell-Derived Cardiomyocytes (hiPSC-CMs) as a Platform to Model Arrhythmogenic Diseases

Pourrier

Fedida

2020

IJMS

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There is a need for improved in vitro models of inherited cardiac diseases to better understand basic cellular and molecular mechanisms and advance drug development. Most of these diseases are associated with arrhythmias, as a result of mutations in ion channel or ion channel-modulatory proteins. Thus far, the electrophysiological phenotype of these mutations has been typically studied using transgenic animal models and heterologous expression systems. Although they have played a major role in advancing the understanding of the pathophysiology of arrhythmogenesis, more physiological and predictive preclinical models are necessary to optimize the treatment strategy for individual patients. Human induced pluripotent stem cell-derived cardiomyocytes (hiPSC-CMs) have generated much interest as an alternative tool to model arrhythmogenic diseases. They provide a unique opportunity to recapitulate the native-like environment required for mutated proteins to reproduce the human cellular disease phenotype. However, it is also important to recognize the limitations of this technology, specifically their fetal electrophysiological phenotype, which differentiates them from adult human myocytes. In this review, we provide an overview of the major inherited arrhythmogenic cardiac diseases modeled using hiPSC-CMs and for which the cellular disease phenotype has been somewhat characterized.

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“…6C). Additionally, our previously published investigations indicated that extended in vitro cell culture may lead to the improvement of cardiomyocyte maturation parameters, such as sarcomere organization, accumulation of cardiac mitochondrial density and transition from alpha to beta MHC isoform 42 .…”

Section: Discussionmentioning

confidence: 98%

Tissue-specific promoter-based reporter system for monitoring cell differentiation from iPSCs to cardiomyocytes

Fiedorowicz

Rozwadowska

Zimna

et al. 2020

Sci Rep

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The possibility of using stem cell-derived cardiomyocytes opens a new platform for modeling cardiac cell differentiation and disease or the development of new drugs. Progress in this field can be accelerated by high-throughput screening (HTS) technology combined with promoter reporter system. The goal of the study was to create and evaluate a responsive promoter reporter system that allows monitoring of iPSC differentiation towards cardiomyocytes. The lentiviral promoter reporter system was based on troponin 2 (TNNT2) and alpha cardiac actin (ACTC) with firefly luciferase and mCherry, respectively. The system was evaluated in two in vitro models. vitro cell culture (confirmed by I-FISH, ddPCR, qPCR). Second, differentiated and contracting control cardiomyocytes (obtained from control non-reporter transduced iPSCs) were subsequently transduced with TNNT2-luc-T2A-Puro-CMV-GFP and hACTC-mcherry-WPRE-EF1-Neo lentiviruses to observe the functionality of obtained cardiomyocytes. Our results indicated that the reporter modified cell lines can be used for HTS applications, but it is essential to monitor the stability of the reporter sequence during extended cell in vitro culture. First, system followed the differentiation of TNNT2-luc-T2A-Puro-mCMV-GFP and hACTC-mcherry-WPRE-EF1-Neo from transduced iPSC line towards cardiomyocytes and revealed the significant decrease in both inserts copy number during the prolonged in

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The impact of in vitro cell culture duration on the maturation of human cardiomyocytes derived from induced pluripotent stem cells of myogenic origin

Cited by 63 publications

References 59 publications

Cardiac Hypertrophy in a Dish: A Human Stem Cell Based Model

Cardiac Hypertrophy in a Dish: A Human Stem Cell Based Model

The Emergence of Human Induced Pluripotent Stem Cell-Derived Cardiomyocytes (hiPSC-CMs) as a Platform to Model Arrhythmogenic Diseases

Tissue-specific promoter-based reporter system for monitoring cell differentiation from iPSCs to cardiomyocytes

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