Emerging technologies and therapeutic strategies have been highlighted that bridge the gap between bench and bedside, demonstrating the translational potential of iPSC-derived 3D cardiac systems for precision medicine in cardiovascular disease.
Abstract
Abstract Cardiomyopathies represent a substantial global health burden, yet progress in developing effective therapies was long constrained by the absence of physiologically relevant human models to understand molecular mechanisms and evaluating interventions. The advent of induced pluripotent stem cell (iPSC) technology has driven major advances in cardiomyopathy research, although important limitations persist. Current iPSC-based systems exhibit incomplete cellular maturation, lack functional vasculature and chamber level architecture, and possess an immature extracellular matrix. These factors must be carefully weighed when interpreting disease phenotypes and drug response data. Induced pluripotent stem cell-derived in vitro three-dimensional (3D) cardiac tissues have significantly addressed these issues, thereby expanding our ability to model human cardiac disease, offering more physiologically relevant platforms for mechanistic studies, drug screening, and translational research. This concise review synthesizes recent advances in the development of iPSC-derived 3D cardiac systems, such as 3D spheroids, engineered heart tissues, and cardiac organoids, their application to modeling various cardiomyopathies including hypertrophic cardiomyopathy, dilated cardiomyopathy, and amyloid-related cardiomyopathies, while discussing critical challenges including cellular maturation, vascularization, and standardization. In this review, emerging technologies and therapeutic strategies have been highlighted that bridge the gap between bench and bedside, demonstrating the translational potential of these models for precision medicine in cardiovascular disease.
Human vascular function depends on tightly coordinated structural, mechanical, and cellular interactions, yet these features remain difficult to recapitulate in vitro. Induced pluripotent stem cells (iPSCs) enable efficient generation of vascular cell types, including endothelial cells, smooth muscle cells, and pericytes, but current systems often lack functional maturity and physiological relevance. Recent advances in vascular organoid engineering provide new opportunities to address this limitation. By integrating self-organization, co-culture, and bioengineering approaches, iPSC-derived systems can form three-dimensional vascular networks with increasing physiological relevance. Emerging evidence from studies of iPSC-derived vascular systems, spanning both two-dimensional differentiation models and three-dimensional organoid platforms, highlights the critical roles of hemodynamic cues, including shear stress and perfusion, together with metabolic and immune signaling, in driving the coordinated maturation of endothelial and mural compartments. These platforms enable modeling of key vascular pathologies, including inflammation, vascular remodeling, and barrier dysfunction, while gene editing further facilitates mechanistic investigation in patient-specific contexts. Together, iPSC-derived vascular systems provide a scalable and physiologically relevant platform for disease modeling, drug discovery, and regenerative medicine.
Chonggui Jiang, Pan Cui, Liyan Gong· Vascular pharmacology· 0 citations
Heart failure is a major clinical and economic burden that afflicts 60 million individuals worldwide. Guideline-directed medical therapies can slow disease progression, but they cannot restore the loss of cardiomyocytes. Over the past two decades, human pluripotent stem cell (hPSC)-based technology has emerged as a leading approach to overcome limited cardiac regenerative capacity, offering a scalable source of functional human cardiomyocytes. The field is now at a pivotal translational stage, as advances in differentiation and tissue engineering have enabled hPSC-based products to enter first-in-human clinical trials. In this review, we summarize the pathophysiological rationale for cell-based therapy in heart failure with reduced ejection fraction. Then, we examine the preclinical foundations of distinct hPSC-derived product formats, including cell suspensions, epicardial sheets, engineered heart muscle, and cardiac spheroids, each with distinct tradeoffs and translational considerations. We conclude by providing updates on ongoing and recently completed clinical trials, evaluating their safety, feasibility, and preliminary efficacy outcomes.
Byron W H Mui, E. Neofytou, Joseph C. Wu· Cell Reports Medicine· 0 citations
Compartment-specific induced pluripotent stem cell (iPSC)-derived cardiomyocytes provide a powerful resource to study cellular and molecular underpinnings of congenital heart disease (CHD) and acquired cardiovascular disease (CVD). Human heart development requires coordinated and complex regulation of key signaling pathways including Notch, BMP, Wnt, Nodal, and Shh during each step of heart morphogenesis. Compartment-specific cardiac cells for modeling the various structures in the heart can be generated by fine tuning these signaling pathways in a sequential manner thereby mimicking spatiotemporal regulation during embryonic heart morphogenesis. In this review, we provide a brief overview of key signaling pathways that are responsible for forming the distinct structures of the heart originating from the first heart field (FHF) and second heart field (SHF). We then summarize recent differentiation protocols that leverage key heart-development related signaling molecules to generate compartment-specific cardiomyocytes and organoids for disease modeling and therapeutic development in cardiovascular disease.
Aaron Argall, Rabina Shrestha, Jiyoon Lee et al.· Cells· 0 citations
Oligonucleotide-based therapeutics represent a rapidly advancing class of drugs with significant potential for treating cardiovascular diseases; however, achieving efficient delivery to cardiac tissue remains a critical and unresolved challenge. A key obstacle is the limited availability of robust, physiologically relevant human in vitro models capable of supporting quantitative assessment of oligonucleotide cellular uptake and intracellular distribution. A detailed, step-by-step protocol is presented for generating self-organizing, 3D cardioids from human induced pluripotent stem cells (iPSCs) and applying them as a platform to evaluate the uptake of fluorescently labeled oligonucleotides. The protocol guides users through directed cardiac differentiation in suspension culture by temporally modulating Wnt/β-catenin signaling, enabling sequential specification of iPSCs through the mesoderm, cardiac mesoderm, and cardiomyocyte progenitor stages. Under these conditions, cells spontaneously self-assemble into beating, cavity-containing three-dimensional structures that express canonical cardiomyocyte markers. The resulting cardioids provide a scalable, experimentally tractable platform for imaging-based assessment of oligonucleotide uptake efficiency, supporting the development and optimization of delivery strategies for cardiac applications.
Ana Da Rosa Soares, Marta Furtado, Rita Tomás et al.· Journal of Visualized Experi...· 0 citations
Transitioning from animal to human cell sources represents a critical milestone in cardiac tissue engineering and biomedical research. Neonatal rat ventricular myocytes (NRVMs) have long served as the functional benchmark for engineered cardiac tissues; however, their rodent origin limits clinical relevance. Human-induced pluripotent stem cell-derived cardiomyocytes (hiPSC-CMs) offer a renewable, species-specific alternative but remain restricted by immature structure and function, small-scale yield, and high batch variability by conventional two-dimensional monolayer (2D-Mono) differentiation. Here, we systematically evaluated hiPSC-CMs generated by 2D-Mono and three-dimensional embryoid-body (3D-EB) differentiation using identical 15-day Wnt-modulated protocols without additional maturation steps. The 3D-EB method yielded 181 × 106 cells per 100 ml, approximately 2.7-fold higher than the 2D-Mono, while maintaining >80% cTnT+ purity and reduced batch variability. Structural analyses revealed improved sarcomeric organization in 3D-EB tissues compared with 2D-Mono, although both remained less organized than NRVMs, while other morphological parameters were comparable between groups. Functionally, 3D-EB tissues exhibited faster calcium conduction (33.7 cm/s), indicating enhanced electrical coupling relative to 2D-Mono. Although contractile performance remained similar between differentiation formats and below NRVM levels, 3D-EB tissues exhibited consistent structural and functional improvement in calcium wave velocity and contractility over time. These results show that, even without external maturation cues, 3D-EB differentiation yields reproducible, scalable, and human-relevant cardiomyocytes.
Yongjun Jang, K. Shani, Anna Clouvel-Gervaiseau et al.· APL Bioengineering· 0 citations
Organoids derived from human pluripotent stem cells (PSCs) have emerged as powerful in vitro models for studying development, disease, and therapeutic responses, yet their lack of functional vasculature limits growth, maturation, and physiological relevance. Early vascularization strategies relied on human umbilical vein endothelial cells, which lack organ-specific identity and introduce donor variability. The field is now undergoing a paradigm shift toward PSC-derived vasculature, which offers patient-specific, and developmentally stage-matched endothelium with PSC-derived organoids. This review summarizes current strategies for organoid vascularization, with emphasis on both human PSC-derived 2D endothelial cells (EC) and 3D blood vessels. Approaches relying on co-aggregation of differentiated ECs with organ-specific populations or external endothelial coating of pre-formed organoids. These improved survival and functional maturation but remain limited in spatial organization and perfusability. The advances have incorporated pre-formed vascular spheroids and iPSC-derived blood vessel organoids, which can be respectively fused with lineage-specific organoids to generate vascularized assembloids to enhance vascular architecture and tissue maturation. This review further highlights engineering the microenvironment to promote the formation of vascular niche, such as hypoxia modulation, transcriptional regulation, signaling transduction, and extracellular matrix engineering. In addition, we discuss the current limitations as well as future directions of vascularized organoids, including the unmet need for developing tissue-specific ECs, improved engraftment following transplantation, and organ-on-a-chip platforms. Collectively, integrating iPSC-derived vasculature within organoids provides a central framework toward physiologically relevant, perfusable tissues and expands the translational utility of organoid technologies for disease modeling and therapeutic development.
