Generation, functional analysis and applications of isogenic three-dimensional self-aggregating cardiac microtissues from human pluripotent stem cells

Campostrini, Giulia; Meraviglia, Viviana; Giacomelli, Elisa; Helden, Ruben W.J. van; Yiangou, Loukia; Davis, Richard P.; Bellin, Milena; Orlova, Valeria V.; Mummery, Christine L.

doi:10.1038/s41596-021-00497-2

Cited by 61 publications

(84 citation statements)

References 81 publications

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“… Giacomelli et al (2017) were able to generate cardiac microtissues by co-culturing purified CMs and ECs at an 85–15% ratio in a restricted culturing space, and show improved maturation and response to drug stimulation. They later show that these microtissues can also be formed including CFs, highlighting the microtissues utility in modelling cell-type specific diseases ( Campostrini et al, 2021 ). These multi-lineage spheroid systems are highly applicable for cardiotoxicity drug screens and disease modelling due to their controlled nature leading to high reproducibility.…”

Section: Current 3d Tissue Models Of the In Vivo Heartmentioning

confidence: 97%

Insights to Heart Development and Cardiac Disease Models Using Pluripotent Stem Cell Derived 3D Organoids

Chan

Soh

2021

Front. Cell Dev. Biol.

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Medical research in the recent years has achieved significant progress due to the increasing prominence of organoid technology. Various developed tissue organoids bridge the limitations of conventional 2D cell culture and animal models by recapitulating in vivo cellular complexity. Current 3D cardiac organoid cultures have shown their utility in modelling key developmental hallmarks of heart organogenesis, but the complexity of the organ demands a more versatile model that can investigate more fundamental parameters, such as structure, organization and compartmentalization of a functioning heart. This review will cover the prominence of cardiac organoids in recent research, unpack current in vitro 3D models of the developing heart and look into the prospect of developing physiologically appropriate cardiac organoids with translational applicability. In addition, we discuss some of the limitations of existing cardiac organoid models in modelling embryonic development of the heart and manifestation of cardiac diseases.

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Section: Current 3d Tissue Models Of the In Vivo Heartmentioning

confidence: 97%

Insights to Heart Development and Cardiac Disease Models Using Pluripotent Stem Cell Derived 3D Organoids

Chan

Soh

2021

Front. Cell Dev. Biol.

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“…The hiPSCs were differentiated into cardiomyocytes as previously described (Campostrini et al, 2021).…”

Section: Differentiation and Culture Of Hipsc-cmsmentioning

confidence: 99%

“…The hiPSC-CMs were cryopreserved at differentiation d20 or d21 as previously described in a freezing medium comprising of 90% Knockout Serum Replacement (Gibco) and 10% DMSO (Brink et al, 2020). Subsequent thawing and seeding of the cells were performed as previously described (Brink et al, 2020;Campostrini et al, 2021).…”

Section: Differentiation and Culture Of Hipsc-cmsmentioning

confidence: 99%

STRAIGHT-IN: A platform for high-throughput targeting of large DNA payloads into human pluripotent stem cells

Grandela

Blanch-Asensio

Brandão

et al. 2021

Preprint

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Inserting large DNA payloads (>10 kb) into specific genomic sites of mammalian cells remains challenging. Applications ranging from synthetic biology to evaluating the pathogenicity of disease-associated variants for precision medicine initiatives would greatly benefit from tools that facilitate this process. Here, we merge the strengths of different classes of site-specific recombinases and combine these with CRISPR/Cas9-mediated homologous recombination to develop a strategy for stringent site-specific replacement of genomic fragments at least 50 kb in size in human induced pluripotent stem cells (hiPSCs). We demonstrate the versatility of STRAIGHT-IN (Serine and Tyrosine Recombinase Assisted Integration of Genes for High-Throughput INvestigation) by: (i) inserting various combinations of fluorescent reporters into hiPSCs to assess excitation-contraction coupling cascade in derivative cardiomyocytes, and; (ii) simultaneously targeting multiple variants associated with inherited cardiac arrhythmic disorder into a pool of hiPSCs. STRAIGHT-IN offers a precise approach to generate genetically-matched panels of hiPSC lines efficiently and cost-effectively.

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“…The hiPSCs were differentiated into cardiomyocytes using the Pluricyte Cardiomyocyte Differentiation kit (Ncardia, Leiden, Netherlands) according to the protocol of the manufacturer. After 20-21 days of differentiation, the hiPSC-CMs were dissociated and cryopreserved as previously described (van den Brink et al, 2020a;Campostrini et al, 2021). For replating, frozen hiPSC-CMs were thawed as described (Brandão et al, 2020) and replated in Medium C (Ncardia) with RevitaCell Supplement (1:100 dilution, Thermo Fisher Scientific).…”

Section: Culture and Differentiation Of Hipscsmentioning

confidence: 99%

The Linkage Phase of the Polymorphism KCNH2-K897T Influences the Electrophysiological Phenotype in hiPSC Models of LQT2

et al. 2021

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While rare mutations in ion channel genes are primarily responsible for inherited cardiac arrhythmias, common genetic variants are also an important contributor to the clinical heterogeneity observed among mutation carriers. The common single nucleotide polymorphism (SNP) KCNH2-K897T is associated with QT interval duration, but its influence on the disease phenotype in patients with long QT syndrome type 2 (LQT2) remains unclear. Human induced pluripotent stem cells (hiPSCs), coupled with advances in gene editing technologies, are proving an invaluable tool for modeling cardiac genetic diseases and identifying variants responsible for variability in disease expressivity. In this study, we have used isogenic hiPSC-derived cardiomyocytes (hiPSC-CMs) to establish the functional consequences of having the KCNH2-K897T SNP in cis- or trans-orientation with LQT2-causing missense variants either within the pore-loop domain (KCNH2A561T/WT) or tail region (KCNH2N996I/WT) of the potassium ion channel, human ether-a-go-go-related gene (hERG). When KCNH2-K897T was on the same allele (cis) as the primary mutation, the hERG channel in hiPSC-CMs exhibited faster activation and deactivation kinetics compared to their trans-oriented counterparts. Consistent with this, hiPSC-CMs with KCNH2-K897T in cis orientation had longer action and field potential durations. Furthermore, there was an increased occurrence of arrhythmic events upon pharmacological blocking of hERG. Collectively, these results indicate that the common polymorphism KCNH2-K897T differs in its influence on LQT2-causing KCNH2 mutations depending on whether it is present in cis or trans. This study corroborates hiPSC-CMs as a powerful platform to investigate the modifying effects of common genetic variants on inherited cardiac arrhythmias and aids in unraveling their contribution to the variable expressivity of these diseases.

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Generation, functional analysis and applications of isogenic three-dimensional self-aggregating cardiac microtissues from human pluripotent stem cells

Cited by 61 publications

References 81 publications

Insights to Heart Development and Cardiac Disease Models Using Pluripotent Stem Cell Derived 3D Organoids

Insights to Heart Development and Cardiac Disease Models Using Pluripotent Stem Cell Derived 3D Organoids

STRAIGHT-IN: A platform for high-throughput targeting of large DNA payloads into human pluripotent stem cells

The Linkage Phase of the Polymorphism KCNH2-K897T Influences the Electrophysiological Phenotype in hiPSC Models of LQT2

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