Evolving from organoid to assembloid with enhanced cellular interactions
Jiarui Liu1, Yitong Shi1, Xianqin Shen1, Wei Zhang2, Xi Wang1,3(), Kai Wang1()
Department of Physiology and Pathophysiology, School of Basic Medical Sciences, State Key Laboratory of Vascular Homeostasis and Remodeling, Clinical Stem Cell Research Center, Peking University Third Hospital, Peking University, Beijing 100191, China
TianXinFu (Beijing) Medical Appliance Co., Ltd., Beijing 102200, China
State Key Laboratory of Female Fertility Promotion, Department of Obstetrics and Gynecology, Peking University Third Hospital, Institute of Advanced Clinical Medicine, Peking University, Beijing 100191, China
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Highlights
• Detailed analysis of the four key assembly strategies for building physiologically relevant assembloids: multi-region, multi-lineage, multi-gradient, and multi-layer.
• Application of assembloids in modeling complex interactions across human systems, including the nervous, digestive, urinary, reproductive, and circulatory systems.
• Identification of key challenges such as reproducibility and vascularization, with proposed solutions using bioengineering techniques and artificial intelligence to improve model accuracy and functionality.
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This review explores the emerging field of assembloid technology, which integrates multiple organoids or cell types into self-organizing three-dimensional (3D) systems to better model inter-tissue communication. It categorizes assembloids into four key assembly strategies and discusses their applications across various human systems, while also addressing challenges and future prospects.
Abstract
In recent years, the study of complex cellular interactions has been hampered by the limitations of traditional two-dimensional (2D) and single-cell type culture systems, which fail to accurately mimic the intricate dynamics of human tissues. To bridge this gap, assembloid technology has emerged as a transformative approach. Assembloids are self-organizing three-dimensional (3D) systems formed by integrating multiple organoids or cell types, providing a more accurate model for studying inter-tissue and inter-organ communication. Here, we categorize current assembloids into four types based on assembly strategies—multi-region, multi-lineage, multi-gradient, and multi-layer—each designed to replicate specific biological phenomena with high fidelity. We also explore the diverse applications of assembloids across various human systems, demonstrating the broad application scope of assembloids. Finally, we highlight the challenges faced by assembloid technology and outline its future prospects. Overall, assembloids represent a powerful platform for advancing research in developmental biology, disease modeling, and drug discovery.
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Comparison of commonly used models in biology.
Multiple factors influence the choice of commonly used models in biology. This figure provides a comparison of the capabilities of commonly used models, including 2D models, conventional scaffold-based 3D models, unguided organoids, region-specific organoids, assembloids, and animal models, in terms of structural complexity, modeling human diseases, reproducibility, drug response predictivity, high-throughput screening compatibility, and ethical concerns. Created with BioRender.com.
Assembly strategies for building the assembloids.
Schematic overview of assembly strategies for the four primary types of assembloids: multi-region, multi-lineage, multi-gradient, and multi-layer. Created with BioRender.com.
Assembloids for recapitulating organ features and disease modelling.
(a) Cortico-motor assembloid integrating human cortical spheroids (hCS), human spinal spheroids (hSpS) and human skeletal muscle spheroids (hSkM)
[
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]. (b) Forebrain assembloid consisting of hCS and human subpallial spheroids (hSS) derived from Timothy Syndrome (TS) individuals
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]. (c) Human neural-perivascular assembloid with pericyte-like cells (PLCs) can be infected with SARS-CoV-2
[
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]. (d) Small cell lung cancer (SCLC)-human cortical organoid (hCO) assembloids mimicked SCLC brain metastasis
[
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]. (e) Murine colon assembloid modelled the interplay between epithelial crypts and stroma
[
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]. (f) Pancreatic ductal adenocarcinoma (PDAC) assembloids can be used in investigating drug resistance mechanisms
[
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]. (g) Bladder assembloids obtained by reorganizing bladder organoids with other components of the microenvironment
[
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]. (h) Locally injured renal assembloids constructed using the multifunctional acoustic tweezer
[
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]. (i) Co-culturing endometrial assembloids containing stromal cells with embryos
[
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]. (j) Human endometrial assembloids with luminal structures are used for modeling endometrial diseases
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,
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]. (k) Combining brain organoids and vascular organoids derived from human pluripotent stem cells to simulate the physiology of the blood–brain barrier and related diseases
[
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]. Created with BioRender.com. CP, cortical plate. SVZ, subventricular zone. VZ, ventricular zone.SARS-CoV-2, Severe Acute Respiratory Syndrome Coronavirus 2. diPDAC. differentiated pancreatic ductal adenocarcinoma. CICs, cancer-initiating cells. iPS cell, induced pluripotent stem cell. ECs, endothelial cells. hPSCs, human pluripotent stem cells. BBB, blood–brain barrier.
Key challenges and future directions in assembloid technology.
Assembloid research faces several critical challenges, includinglow reproducibility, intrinsic heterogeneity, high labor intensity,difficulty in maintaining stability, and lack of vascularization. Toaddress these, standardized quality control and bioengineeringapproaches can improve reproducibility and reduce heterogeneity;artificial intelligence may alleviate labor demands by minimizingmanual intervention; and the development of universal mediacould enhance the long-term stability of assembloids.Additionally, as assembloid technologies progress, ethicalconsiderations must not be overlooked.
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