Three-dimensional cardiac organoid formation accelerates the functional maturation of human induced pluripotent stem cell-derived cardiomyocytes

Lee, Hanbyeol; Im, Jeong Suk; Choi, Da Bin; An, Jieun; Kim, Su-Bin; Yeon, Seunghee; Yoon, Seulgi; Woo, Dong-Hun

doi:10.51335/organoid.2022.2.e14

Cited by 5 publications

(4 citation statements)

References 56 publications

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“…We previously generated hiPSC-derived cardiac organoids (COs), and the CO formation resulted in the enhanced maturity of hiPSC-derived cardiomyocytes [ 18 ]. However, these COs could not mimic the diverse cellular composition of the human heart.…”

Section: Resultsmentioning

confidence: 99%

“…The hiPSCs were expanded and cultured under feeder-free conditions on Matrigel (Corning, USA) coated T75 tissue culture flasks in the presence of mTeSR1 (Stem Cell Technologies, Canada). Cardiac organoid (CO) differentiation was performed as described in our previous reports [ 18 ]. To differentiate heart organoids (HOs), we initially applied the differentiation procedures used for COs to induce mesodermal differentiation.…”

Section: Methodsmentioning

confidence: 99%

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Modeling acute myocardial infarction and cardiac fibrosis using human induced pluripotent stem cell-derived multi-cellular heart organoids

Song,

Choi,

et al. 2024

Cell Death Dis

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Heart disease involves irreversible myocardial injury that leads to high morbidity and mortality rates. Numerous cell-based cardiac in vitro models have been proposed as complementary approaches to non-clinical animal research. However, most of these approaches struggle to accurately replicate adult human heart conditions, such as myocardial infarction and ventricular remodeling pathology. The intricate interplay between various cell types within the adult heart, including cardiomyocytes, fibroblasts, and endothelial cells, contributes to the complexity of most heart diseases. Consequently, the mechanisms behind heart disease induction cannot be attributed to a single-cell type. Thus, the use of multi-cellular models becomes essential for creating clinically relevant in vitro cell models. This study focuses on generating self-organizing heart organoids (HOs) using human-induced pluripotent stem cells (hiPSCs). These organoids consist of cardiomyocytes, fibroblasts, and endothelial cells, mimicking the cellular composition of the human heart. The multi-cellular composition of HOs was confirmed through various techniques, including immunohistochemistry, flow cytometry, q-PCR, and single-cell RNA sequencing. Subsequently, HOs were subjected to hypoxia-induced ischemia and ischemia-reperfusion (IR) injuries within controlled culture conditions. The resulting phenotypes resembled those of acute myocardial infarction (AMI), characterized by cardiac cell death, biomarker secretion, functional deficits, alterations in calcium ion handling, and changes in beating properties. Additionally, the HOs subjected to IR efficiently exhibited cardiac fibrosis, displaying collagen deposition, disrupted calcium ion handling, and electrophysiological anomalies that emulate heart disease. These findings hold significant implications for the advancement of in vivo-like 3D heart and disease modeling. These disease models present a promising alternative to animal experimentation for studying cardiac diseases, and they also serve as a platform for drug screening to identify potential therapeutic targets.

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Section: Resultsmentioning

confidence: 99%

Section: Methodsmentioning

confidence: 99%

Modeling acute myocardial infarction and cardiac fibrosis using human induced pluripotent stem cell-derived multi-cellular heart organoids

Song,

Choi,

et al. 2024

Cell Death Dis

Self Cite

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“…The hPSCs cardiomyocytes were grown in a 3D organotypic structure, promoting more mature ion channel expression and morphological features [58]. To take advantage of these properties, cardiomyocytes were first grown in a 3D cardioid system according to the protocol in [6].…”

Section: Cellsmentioning

confidence: 99%

Nanoimprinted Hierarchical Micro-/Nanostructured Substrates for the Growth of Cardiomyocyte Fibers

Mühlberger,

Kopp,

Deyett

et al. 2023

Nanomanufacturing

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Investigating the behavior of cardiomyocytes is an important part of drug development. We present a structure and a related nanoimprint-based fabrication method, where the cardiomyocytes form isolated fibers, which is beneficial for drug testing, more closely representing the structure of the cardiomyocytes in vivo. We found that channel structures with walls with a rough top surface stimulate cardiomyocytes to form such fibers, as desired. Nanoimprint lithography is used as a fast and cost-efficient method to fabricate our hierarchically structured cell growth substrates.

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“…Human induced pluripotent stem cells (hiPSCs) obtained from NEXEL Co., Ltd. were used as the cellular source for generating 3D cardiac organoids. The detailed method for generating hiPSCs was previously described [25]. The hiPSC cells were cultivated and maintained on Growth Factor Reduced (GFR) Matrigel® (356231, Corning)-coated plates in DMEM/F12 (11320033, GibcoTM) at a 1:100 dilution with mTeSRTM1 (85851, StemCellTM Technologies) medium containing 0.5% penicillin/streptomycin (0503; Thermo Fisher Scienti c).…”

Section: Culture and Expansion Of Hipscsmentioning

confidence: 99%

Assessment of the Optimal Generation Period and Size of Human iPSC-Derived Cardiac Organoids for Cardiotoxicity Drug Testing

Lee

Song

Woo

et al. 2023

Preprint

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Background Human-induced pluripotent stem cell-derived cardiac organoids (hiPSC-COs) have emerged as valuable tools for cardiotoxicity drug screening, given their ability to recapitulate in vivo cardiac tissue functions and facilitate rapid assessment of drug stability to prevent toxicity. However, the lack of established criteria for the differentiation period and size during the generation of functional hiPSC-COs can introduce low accuracy in drug screening responses results. Hence, it is crucial to establish appropriate criteria for the generation period and size of hiPSC-COs to ensure reliable cardiotoxicity drug screening. Methods In this study, we generated different-sized hiPSC-COs in two types of microwell arrays through a one-stop generation method. The two-sized hiPSC-COs were continuously monitored until a stable cardiac beating rate was confirmed during the differentiation period. We evaluated and compared the functionality such as calcium transients at the selected differentiation day that showed a stable beating rate with a specific focus on determining the minimal differentiation period required for generating functional hiPSC-COs. A physiological test was conducted to verify the reactivity to the drug in hiPSC-COs according to the differentiation period and size. Subsequently, we conducted a cardiotoxicity drug screening test using compounds known to induce in vivo heart failure. Finally, characterization was analyzed by immunostaining assay to compare and confirm the phenotype of the two-sized hiPSC-COs at the selected differentiation period. Results During the differentiation period to generate hiPSC-COs, we identified the time point at which the smaller organoids among the two sizes of hiPSC-COs began to show a stable beating rate, which was an optimal period to lead to meaningful response results to cardiotoxicity drugs. Moreover, large organoids confirmed that cardiac properties disappeared as the differentiation period progressed, suggesting insight into the size limitation on the generation of hiPSC-COs for cardiotoxicity testing. Furthermore, an additional analysis method was proposed for subtle reactions that are difficult to confirm solely using the beating rate analysis in drug response testing. Conclusion We expect that these findings may contribute to the field of drug development by ensuring significant drug response results and enhancing the reliability of cardiotoxicity testing using hiPSC-COs.

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Three-dimensional cardiac organoid formation accelerates the functional maturation of human induced pluripotent stem cell-derived cardiomyocytes

Cited by 5 publications

References 56 publications

Modeling acute myocardial infarction and cardiac fibrosis using human induced pluripotent stem cell-derived multi-cellular heart organoids

Modeling acute myocardial infarction and cardiac fibrosis using human induced pluripotent stem cell-derived multi-cellular heart organoids

Nanoimprinted Hierarchical Micro-/Nanostructured Substrates for the Growth of Cardiomyocyte Fibers

Assessment of the Optimal Generation Period and Size of Human iPSC-Derived Cardiac Organoids for Cardiotoxicity Drug Testing

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