Modeling the human limb skeletal development using human pluripotent stem cell-derived skeletal assembloid

bioRxiv (Cold Spring Harbor Laboratory), Journal Year: 2025, Volume and Issue: unknown

Published: Jan. 10, 2025

Despite recent advances in pluripotent stem cell-based approaches to induce skeletal cells, recapitulating human limb development terms of structure and longitudinally oriented growth remains an unresolved challenge. Here, we report a method differentiate cells into region-specific organoids harboring GDF5+PRG4+ interzone/articular chondrocyte progenitors (IZ/ACPs) SP7+ plate chondrocytes (GPCs) via PRRX1⁺ limb-bud mesenchymal cells. Comparative analysis demonstrated marked similarities IZ/ACP GPC the embryonic limb, graft fate regenerative capacity vivo were further characterized. We also mimicked developmental process spatially structured manner by vertically positioning two at both ends organoid generate assembloid. Notably, this assembloid recapitulated endochondral ossification with longitudinal upon transplantation. In summary, our study provides novel research platform for disease.

Language: Английский

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Multi-Region Brain Organoid: Fusion Organoid with Cerebral, Endothelial and Mid-Hindbrain Components

bioRxiv (Cold Spring Harbor Laboratory), Journal Year: 2025, Volume and Issue: unknown

Published: Jan. 21, 2025

Abstract Brain organoid technology has revolutionized our ability to model human neurodevelopment in vitro. However, current techniques remain limited by their reliance on simplified endothelial cell populations rather than a complete system. We engineered Multi-Region Organoids (MRBOs) that integrate cerebral, mid/hindbrain, and complex organoids into one structure. Different from the earlier approaches based isolated HUVECs, contain diverse vascular types, including progenitors, mature cells, pericytes, proliferating angiogenic cells stromal cells. Our strategy employs sequential modulation of key developmental pathways generate individual organoids, followed optimized fusion conditions maintain regional identities while supporting cellular integration. Single-nucleus RNA sequencing shows MRBOs develop discrete neural specific each brain region alongside specialized establish paracrine signalling networks. Integration analysis with fetal data contribute 80% clusters found tissue (Carnegie stages 12-16), whereas CellChat identifies 13 previously uncharacterized endothelial-neural interactions. Notably, we uncover endothelial-derived factors support persistence intermediate progenitor during hindbrain development, but not cerebral thereby revealing new role for patterning. This platform enables matching multiple regions, fully incorporating nature at same time providing opportunities studying neurodevelopmental disorders which neural-endothelial interactions are disrupted. MRBO system establishes foundation investigating processes, an enabling context closer physiological relevance.

Language: Английский

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Developing advanced organoids: challenges, progress, and outlook

BioTechniques, Journal Year: 2025, Volume and Issue: unknown, P. 1 - 6

Published: Jan. 29, 2025

Organoids, self-organizing 3D structures created from a variety of cell sources, offer unique advantages for studying organ development, modeling diseases, discovering new drugs, and creating regenerative therapies. However, their ability to completely mimic complex in vivo structure function has been hindered by the lack all relevant types found each organ; heterogeneity between organoids; variable reproducibility; mature phenotype; integrated neural, vascular, hematopoietic networks. To address these critical challenges, various strategies are being rapidly advanced include co-culturing co-differentiating multiple create region-and lineage-specific organoids together, including with vascular organoids, assembloids; using organoid-on-a-chip technology integrate perfusable vasculature within bioprinting organoids. This brief overview explores how converging disciplines stem biology, developmental bioengineering technologies have progressed creation increasingly sophisticated organoid models, provides an outlook on remaining challenges might be addressed.

Language: Английский

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Organoids from pluripotent stem cells and human tissues: When two cultures meet each other

Developmental Cell, Journal Year: 2025, Volume and Issue: 60(4), P. 493 - 511

Published: Feb. 1, 2025

Language: Английский

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The power of mouse models in the diagnostic odyssey of patients with rare congenital anomalies

Mammalian Genome, Journal Year: 2025, Volume and Issue: unknown

Published: March 18, 2025

Abstract Congenital anomalies are structural or functional abnormalities present at birth, which can be caused by genetic environmental influences. The availability of genome sequencing has significantly increased our understanding congenital anomalies, but linking variant identification to relevance and definitive diagnosis remains challenging. Many genes have unknown poorly understood functions, with a lack clear genotype-to-phenotype correlations, it difficult move from discovery diagnosis. Thus, for most there still exists “diagnostic odyssey” presents significant burden patients, families society. Animal models essential in the gene process because they allow researchers validate candidate function disease progression within intact organisms. However, use advanced model systems continues limited due complexity efficiently generating clinically relevant animals. Here we focus on precisely engineered mice variant-to-function studies resolving molecular diagnoses creating powerful preclinical covering advances genomics, editing phenotyping approaches as well necessity future initiatives aligning animal modelling deep patient multimodal datasets.

Language: Английский

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Advances in humanoid organoid-based research on inter-organ communications during cardiac organogenesis and cardiovascular diseases

Journal of Translational Medicine, Journal Year: 2025, Volume and Issue: 23(1)

Published: March 28, 2025

The intimate correlation between cardiovascular diseases and other organ pathologies, such as metabolic kidney diseases, underscores the intricate interactions among these organs. Understanding inter-organ communications is crucial for developing more precise drugs effective treatments systemic diseases. While animal models have traditionally been pivotal in studying interactions, human-induced pluripotent stem cells (hiPSCs) offer distinct advantages when constructing vitro models. Beyond conventional two-dimensional co-culture model, hiPSC-derived humanoid organoids emerged a substantial advancement, capable of replicating essential structural functional attributes internal organs vitro. This breakthrough has spurred development multilineage organoids, assembloids, organoids-on-a-chip technologies, which allow enhanced physiological relevance. These technologies shown great potential mimicking coordinated organogenesis, exploring disease pathogenesis, facilitating drug discovery. As central system, heart serves focal point an extensively studied network interactions. review focuses on advancements challenges organs, presenting comprehensive exploration this cutting-edge approach research.

Language: Английский

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Cellular interplay in brain organoids: Connecting cell-autonomous and non-cell-autonomous mechanisms in neurodevelopmental disease

Current Opinion in Neurobiology, Journal Year: 2025, Volume and Issue: 92, P. 103018 - 103018

Published: April 11, 2025

The field of brain organoids has experienced a period rapid and transformative growth, enabling researchers to investigate complex human biological mechanisms that were previously deemed intractable. This review provides an overview the current landscape organoids, with particular focus on their relevance in context neurodevelopmental disorders. It also emphasizes crucial role these models play elucidating both cell-autonomous non-cell-autonomous mechanisms. We describe how two mechanisms, often considered be independent, are intricately interconnected. In conclusion, this aims highlight utilization considerably advanced our comprehension disorders, while delineating prospective avenues for investigating conditions.

Language: Английский

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Current Development of iPSC-Based Modeling in Neurodegenerative Diseases

International Journal of Molecular Sciences, Journal Year: 2025, Volume and Issue: 26(8), P. 3774 - 3774

Published: April 16, 2025

Over the past two decades, significant advancements have been made in induced pluripotent stem cell (iPSC) technology. These developments enabled broader application of iPSCs neuroscience, improved our understanding disease pathogenesis, and advanced investigation therapeutic targets methods. Specifically, optimizations reprogramming protocols, coupled with neuronal differentiation maturation techniques, greatly facilitated generation iPSC-derived neural cells. The integration cerebral organoid technology CRISPR/Cas9 genome editing has further propelled neurodegenerative diseases to a new stage. Patient-derived or CRISPR-edited neurons organoids now serve as ideal models, contributing pathophysiology identifying novel candidates. In this review, we examine development iPSC-based models diseases, including Alzheimer’s disease, Parkinson’s Huntington’s disease.

Language: Английский

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Chimeric brain models: Unlocking insights into human neural development, aging, diseases, and cell therapies

Neuron, Journal Year: 2025, Volume and Issue: unknown

Published: April 1, 2025

Language: Английский

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Bioengineering innovations for neural organoids with enhanced fidelity and function

Cell stem cell, Journal Year: 2025, Volume and Issue: 32(5), P. 689 - 709

Published: May 1, 2025

Language: Английский

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