Landscape of human organoids: Ideal model in clinics and research.

Organoid technology has played a pivotal role in the advancement and evaluation of new pharmaceuticals. While organoids are not directly involved in the creation of novel drugs, they are essential for facilitating the discovery of promising drug candidates via drug screening and testing. 162 , 163 Specifically, organoids derived from colorectal cancer patients have been instrumental in assessing responses to inhibitors of tankyrases, enzymes that are crucial in Wnt signaling and are frequently disrupted in colorectal cancer. This testing has led to the identification of new drug candidates targeting the Wnt pathway. 68 Moreover, organoids from breast and lung cancer patients with certain mutations have been valuable in predicting sensitivity to PARP inhibitors, drugs that focus on DNA-repair mechanisms, 68 , 164 , 165 contributing significantly to the development of individualized treatment plans involving PARP inhibitors. An additional study revealed that organoids possessing ARID1A missense mutations exhibited particular sensitivity to ATR inhibitors, which affect the DNA-damage response. This discovery holds significant promise for the development of new ATR-targeting drugs for cancers with ARID1A mutations. 166 Furthermore, organoids from endometrial and metastatic tumors responded to STAT3 inhibitors, hinting at the potential for novel drugs that target STAT3 signaling in endometrial adenocarcinoma. 167 Organoid lines from mesothelioma patients retained biomarkers and histological features of the patients’ tumors. 168 In drug testing with cisplatin and pemetrexed, the organoids mirrored the patients’ responses, demonstrating the potential of organoids in predicting treatment outcomes for mesothelioma. Finally, organoids from appendiceal carcinoma were used in immunotherapy trials, paving the way for the creation of new immunotherapeutic agents tailored to the unique characteristics of appendiceal tumors. 167 In every facet of pharmaceutical development, organoids manifest indispensable significance, providing robust support and verification from foundational research to clinical trials and all the way to the guidance of personalized medication, facilitating all-encompassing success in drug innovation. The emergence of organoids in high-throughput drug screening has transformed drug discovery by enabling the targeted identification of effective molecules from large compound libraries. 162 , 163 Recent advances in automation and systematic platforms have made organoids a gold standard in drug screening with the potential to streamline drug-discovery pipelines. 169 , 170 , 171 Their ability to efficiently identify the most effective molecules from large compound libraries is transformative, and organoids are poised to become an integral part of drug-discovery pipelines. A large-scale organoid-based screening platform was used to test 1,172 Food and Drug Administration-approved compounds on pancreatic cancer organoids, and 22 drugs with anticancer effects were identified. This effort underscored the importance of three-dimensional culture-based platforms for drug screening, as some drugs that were effective in organoids did not show the same effects in two-dimensional cultures. 166 One of the most critical aspects of drug development is the early identification of toxicological profiles. Traditional animal models often fail to capture the full spectrum of human-specific toxicities, leading to costly failures in late-stage clinical trials. Organoids, because of their human-derived cells and tissue-specific architectures, constitute a more accurate model for assessing drug toxicity. 172 , 173 Kidney organoids, 174 , 175 for instance, have been used to evaluate nephrotoxicity, which is a significant concern in human pharmacology. Organoids are crucial and powerful tools not only for testing compound efficacy and toxicity but also for understanding disease mechanisms and validating therapeutic targets. 176 In cancer research, patient-derived tumor organoids play an important role in validating oncogenes and tumor-suppressor genes, thereby offering insights into drug mechanisms. In a study by Hirt et al. 177 reported by Mainardi and Bernards, 166 a newly derived pancreatic organoid biobank comprising 31 three-dimensional cell lines derived from tumors and nine from healthy tissue was used to investigate novel drug-gene interactions with potential translational value. The evaluation of drug responses in a context that mimics human physiology is crucial for the success of any therapeutic intervention. The advantages of organoids in drug responses can even be applied to clinical trials of new drugs. 81 , 178 Clinical trials are the bottleneck of drug development, with significant time and resource investments followed by high failure rates. Organoids have the potential to increase the success rate of clinical trials by providing a more accurate representation of patients’ diseased tissue and predicting their responses to specific therapies. 179 Organoids derived from patients enrolled in a trial could be used in parallel to evaluate drug responses, helping to adapt or modify the trial’s course in real time. This personalized approach to clinical trials holds great promise for improving patient outcomes and reducing the time and cost associated with drug development. 180 The aim of personalized medicine is to tailor treatments to individual patients based on their genetic and physiological characteristics. In oncology, patient-derived tumor organoids can be used to screen for effective chemotherapy agents, thereby personalizing treatment plans. 180 , 181 , 182 , 183 The ability to evaluate drug responses in patient-specific organoids holds great promise for improving treatment outcomes and reducing adverse effects. Immunotherapies have revolutionized cancer treatment, but their efficacy varies among patients. Organoids constitute a powerful platform for studying the immune microenvironment and developing novel immunotherapies. 184 , 185 Researchers can assess the efficacy of immunotherapeutic treatments by coculturing organoids with immune cells, such as tumor-infiltrating lymphocytes. 184 The study of tumor-infiltrating lymphocytes within organoids has also introduced new avenues for personalized immunotherapies tailored to the specific immunological landscape of each patient’s tumor. 186 , 187 The ability to model complex diseases ex vivo has been significantly advanced by the advent of organoid technology and gene-editing techniques. For instance, CRISPR-Cas9 has been instrumental in creating lung cancer models by introducing specific mutations into lung organoids. 188 Such models offer a multifaceted approach to understanding complex diseases and have therefore been particularly useful in studying developmental and genetic diseases ( Figures 6 A and 6B). Figure 6 The organoid epoch: A paradigm shift in drug development (A) Organoid technology: advancing drug development across all stages. Organoids are crucial in influencing the drug-development sector, providing extensive assistance throughout the entire process. (B) The various applications of organoid models in biological research. Organoids play a pivotal role in the drug-development industry, offering a wide range of applications that streamline and enhance the process. (C) Comparing the cost and time efficiency of drug development: traditional methods with organoid-based approaches. Organoids represent a fundamental change in medication development, providing notable benefits compared to conventional approaches. The organoid epoch: A paradigm shift in drug development (A) Organoid technology: advancing drug development across all stages. Organoids are crucial in influencing the drug-development sector, providing extensive assistance throughout the entire process. (B) The various applications of organoid models in biological research. Organoids play a pivotal role in the drug-development industry, offering a wide range of applications that streamline and enhance the process. (C) Comparing the cost and time efficiency of drug development: traditional methods with organoid-based approaches. Organoids represent a fundamental change in medication development, providing notable benefits compared to conventional approaches. Ex vivo modeling of complex diseases in organoids, aided by advanced gene-editing techniques, represents a cost-effective and ethical alternative to traditional drug-development methods. Organoids accurately replicate human physiology, thereby reducing the time and costs associated with drug discovery ( Figure 6 C). 189 Their utility in academic research and clinical application is significant, offering the potential to revolutionize the field and pave the way for more efficient therapeutic advancements.

Organoid research, originating in the early 20th century, explores the self-organizing properties of cells ( Figure 1 A). The initial studies involved cultivating mechanically dissociated sponge cells to create functional organisms in vitro . H.V. Wilson’s observations in 1907 marked the inception of organoid research, revealing the self-organization and regeneration of dissociated sponge cells. 9 , 10 , 11 In subsequent decades, several groups performed dissociation-reaggregation experiments to generate different types of organs from dissociated amphibian pronephros and chick embryos, 12 , 13 , 14 demonstrating the ability of cells to self-organize and form organ-like structures in vitro . Figure 1 Unlocking organoids: A tale of historical discoveries and advanced taxonomies (A) The chronological history of the development of organoids. (B) Numerous novel applications of organoids have surfaced in the realm of life science research in recent years. (C) Further refinement is required in classification as a result of the diversification of organoid sources. Unlocking organoids: A tale of historical discoveries and advanced taxonomies (A) The chronological history of the development of organoids. (B) Numerous novel applications of organoids have surfaced in the realm of life science research in recent years. (C) Further refinement is required in classification as a result of the diversification of organoid sources. In 1981, pluripotent stem cells (PSCs) were isolated from mouse embryos, a milestone in stem cell research. 15 , 16 , 17 , 18 In November 1998, human embryonic stem cells (ESCs) were derived from blastocysts, a breakthrough demonstrating human PSC derivation. 19 The development of human induced pluripotent stem cell (iPSC) technology further revolutionized the field. 20 , 21 The modern era of organoid technology truly began in the 2000s with the development of iPSCs by Shinya Yamanaka’s team at Kyoto University 22 , 23 , 24 , 25 : their pioneering work showed that mouse fibroblasts could be reprogrammed into a pluripotent state by introducing four transcription factors (Oct4, Sox2, Klf4, and c-Myc), generating the first iPSCs. 18 , 22 In 2009, Hans Clevers introduced the term “organoid,” emphasizing self-organization, particularly in cancer research. 26 The term “organoid” draws inspiration from the ancient Greek lexicon, where “organon” is the origin of “organ” and the suffix “-oid” denotes resemblance or likeness. Therefore, “organoid” refers to something that resembles an organ. Researchers such as Lancaster and Knoblich expanded upon this seminal work. 27 They opined that the true essence of organoids lies not solely in their structural replication but in their ability to mirror the cellular diversity, specific functionalities, and even intricate cellular arrangements of the organs they represent. 28 Subsequently, the field of organoid technology advanced rapidly, ushering in a new era in scientific innovation. 29 , 30 Hans Clevers’ intestinal organoids paved the way for new research on intestinal biology and disease mechanisms 31 and continue to be a valuable resource for investigating the molecular mechanisms underlying intestinal development and disease progression. 26 , 32 Cerebral organoids, created by Lancaster et al. in 2013, mimic human brain development and aid in understanding developmental brain disorders. 33 In 2014, groundbreaking advances occurred in prostate organoid research, providing a robust platform for studying prostate cancer dynamics and therapeutic drug testing. 34 , 35 In 2015, kidney organoids derived from iPSCs provided insights into kidney development and disease modeling. 36 , 37 Subsequent years saw the burgeoning of organoid technology across various organ systems, including the pancreas. The development of pancreatic cancer organoids has illuminated the pathophysiological mechanisms that underlie pancreatic ductal adenocarcinoma, fostering advances in personalized medicine and targeted therapeutic strategies. 38 , 39 A surge in methodological refinements in retinal organoid cultivation have occurred since 2018. These organoids recapitulate the functionality of retinal cells, representing a significant leap forward from earlier models that lacked structural and functional fidelity. 40 , 41 Most recently, cardiac organoids emerged, aiding in modeling heart diseases. 42 , 43 Organoids have revolutionized biomedical research, enhancing drug assessments and personalized medicine. 44 In oncology, organoids have provided critical insights into the tumor immune microenvironment, propelling advancements in immunotherapy. 45 These three-dimensional cultures serve as pivotal tools in deciphering the complexities of organ and embryonic development, spotlighting essential elements of morphogenesis and the inception of life. 46 Moreover, the establishment of extensive organoid biobanks has supported personalized research approaches, offering a plethora of tissue-specific samples for in-depth study. In neuroimmune research, organoids have been instrumental in elucidating the intricate interplay between the nervous and immune systems, leading to new insights into a myriad of associated disorders. 47 , 48 The remarkable ability of organoids to replicate organ-level functions within controlled environments is responsible for their indispensable role in precision medicine, disease modeling, and regenerative medicine, enabling a transformative phase in modern medical research ( Figure 1 B). The field of organoid development utilizes a variety of stem cell sources, each with unique characteristics. Among these are the well-known ESCs and the revolutionary iPSCs, both of which fall under the broader category of PSCs ( Figure 1 C). The versatile mesenchymal stem cells also play a crucial role in this domain. 49 , 50 , 51 , 52 , 53 , 54 , 55 Additionally, the world of clinical medicine offers a treasure trove of sources in the form of tissue specimens, both from individuals battling diseases and from healthy donors. 56 , 57 This range includes surgical specimens and other less obvious sources such as nasopharyngeal swabs, which have recently gained prominence. 58 In conclusion, organoid research has introduced a new era in biomedical sciences, enhancing our understanding of life, disease, and therapies.

The field of biomedical research has achieved a significant leap forward with the advent of organoid technology. Their versatility and adaptability make them a valuable tool for a wide range of research applications, with the potential to reshape therapeutic approaches to a range of clinical challenges ( Figure 7 A). The promise of organoids is particularly palpable in the realm of diabetes treatment. Researchers are abuzz regarding the creation of islet-like organoids that respond to glucose, which herald a potential paradigm shift in diabetes management. 190 These organoids house a diverse array of cells: insulin-producing β cells, glucagon-producing α cells, somatostatin-producing δ cells, and pancreatic polypeptide-producing γ cells. 191 Their efficacy was tested by transplantation into diabetic mice, 192 where these organoids mimicked native pancreatic β cells, releasing insulin in response to blood glucose fluctuations and stabilizing glucose levels. Despite the promise of this approach, challenges remain, such as scaling up organoid production and maintaining their long-term functionality. Figure 7 The multifaceted impact of organoid technology in biomedical science (A) The domain of biomedical research has experienced a substantial advancement with the introduction of organoid technology. The valuable attributes of versatility and adaptability possessed by these tools render them highly useful in a diverse array of research applications. Consequently, they hold great potential for revolutionizing therapeutic strategies aimed at addressing a wide range of clinical difficulties. (B) The application of organoids has greatly expanded the scope of disease inquiry. Technological advancements have made it possible to explore scientific realms that were previously inaccessible, especially when with regard to situations that are unique to humans. The multifaceted impact of organoid technology in biomedical science (A) The domain of biomedical research has experienced a substantial advancement with the introduction of organoid technology. The valuable attributes of versatility and adaptability possessed by these tools render them highly useful in a diverse array of research applications. Consequently, they hold great potential for revolutionizing therapeutic strategies aimed at addressing a wide range of clinical difficulties. (B) The application of organoids has greatly expanded the scope of disease inquiry. Technological advancements have made it possible to explore scientific realms that were previously inaccessible, especially when with regard to situations that are unique to humans. Regenerative medicine, which is synonymous with restoration and healing, has found in organoids a robust and versatile ally. 125 These structures are pioneering advances in tissue repair, 50 , 53 , 193 , 194 transplantation, and overall regenerative strategies. 53 In inflammatory bowel disease research, organoids are providing insights into tissue-regeneration mechanisms, paving the way for innovative treatments. 125 Organoid transplantation is also presenting new possibilities. A prime example is the transplantation of human intestinal organoids into mice, 195 which resulted not only in the successful integration of the organoids but also in physiological improvements in the recipient mice. Similarly, research into human kidney organoids transplanted into rats has shown promising results, 196 , 197 including enhanced vascularization and function. Such studies hint at a future in which organ transplant waiting lists could be significantly reduced, with organoids serving as a viable alternative. In biobanking, organoids are revolutionizing the collection, preservation, and utilization of normal and pathological specimens, improving disease modeling and drug screening. 198 Organoids also enable the preservation and study of viruses or microorganisms that are difficult to culture in traditional cell lines, providing more accurate models for research on tropism and immunity induction. 199 , 200 , 201 The intersection of organoids and infectious disease research is also particularly compelling. Organoids offer a dynamic platform for studying host-pathogen interactions, paving the way for deeper insights and innovative treatments. 202 , 203 , 204 For instance, organoids have been pivotal in the study of respiratory viruses. 205 Their application in research related to influenza and respiratory syncytial virus is just the tip of the iceberg. 204 However, research on SARS-CoV-2 is where the potential of organoids is truly showcased. 206 , 207 , 208 , 209 Researchers have used lung and intestinal organoids to gain a nuanced understanding of how the virus invades human cells. 210 , 211 , 212 , 213 These studies are revealing vital data about viral entry, replication, and spread. Furthermore, organoids provide a platform for testing potential drugs against the virus, accelerating the drug-discovery process. 214 , 215 Beyond viral infection, organoids have a role to play in bacterial and parasitic infection research. 202 , 216 Their three-dimensional structure, mirroring that of human tissue, offers a unique environment to study the dynamics of bacterial infections at a cellular level. Additionally, in parasitic research, organoids are illuminating host-helminth interactions, 217 unveiling the intricate relationships between parasites and their human hosts. Moreover, organoid models are a powerful tool for studying female fertility and reproductive biology. These organoids, derived from tissues in the female reproductive tract, enable research on normal reproductive processes and pathologies such as endometriosis, endometrial cancer, and ovarian cancer. 156 , 218 , 219 , 220 Ovarian organoids facilitate the investigation of oocyte growth and maturation along with hormonal stimulation responses. 220 Fallopian tube, endometrial, and cervical organoids are utilized to study various aspects of female reproductive health, including the development and function of the fallopian tubes, the menstrual cycle, endometriosis, endometrial hyperplasia, cervical clear cell carcinoma, and the impact of infections such as Chlamydia trachomatis and human papillomavirus. Additionally, organoids derived from pathological tissues have aided in understanding reproductive disorders and have been valuable for drug screening. 156 , 218 , 220 Organoids also model early developmental events, such as implantation, and can be used to investigate interactions between different cell types such as trophoblast cells and the maternal decidua. Recent studies have also involved the development and characterization of trophoblast organoids and decidual organoids from human placental samples, which serve as models to study the maternal-fetal interface. 221 These organoids have been used to investigate innate immune signaling and the differential antiviral defenses present at the maternal-fetal interface. Moreover, organoids have been employed to study the effects of environmental toxicants on female fertility and reproductive health. 219 They offer a physiologically relevant and controlled system for the examination of intricate cellular processes within a contextually appropriate environment, offering the potential to revolutionize the field of reproductive medicine. Duchenne muscular dystrophy, a debilitating genetic disorder, is also the subject of promising advances due to organoids. Using gene-editing technologies, researchers have been able to correct genetic mutations in iPSC-derived muscle cells, which constitutes a monumental advance in the development of effective treatments for Duchenne muscular dystrophy ( Figure 7 B). 82 , 83 Organoid-based research has also enabled the development of innovative therapeutic strategies for retinal diseases, which have historically been challenging to treat. 84 , 85 Leveraging organoids, scientists have been able to explore the potential of gene augmentation therapy using systems such as CRISPR-Cas9. One notable success has been the promise shown in treating Leber congenital amaurosis, a severe form of inherited blindness: targeted gene therapies have demonstrated potential in restoring vision. 83 In summary, the versatile applications of organoids are undeniably setting the stage for groundbreaking advancement in biomedical research. As we continue to harness their potential, the bridge between laboratory-based discoveries and real-world clinical applications is becoming more robust. The future promises a new era of personalized medicine with organoids at its core, offering hope for myriad clinical challenges.

The advent of organoid technology marks a watershed in biomedical research, transcending previous boundaries of scientific inquiry. This innovation has revolutionized our understanding of human biology and introduced unprecedented avenues in disease management, addressing obstacles once considered insurmountable. At the cusp of this burgeoning field, the possibilities seem limitless, with organoid technology poised to reshape healthcare, research methodologies, and ethical considerations concerning human life. Throughout this review, we have highlighted the multifaceted impact of organoids, emphasizing their utility in personalized medicine, drug discovery, and disease modeling. The bespoke nature of organoid systems, particularly patient-derived organoids, has introduced a new era of precision in biomedical research. By closely mimicking the intricate architecture and functionality of human organs, organoids offer a more representative model for human physiology and pathology, surpassing traditional in vitro and animal models. The capacity of organoids to replicate complex biological processes, such as the tumor immune microenvironment and host-pathogen interactions, positions them as a vanguard in further elucidating disease mechanisms and therapeutic responses. Looking to the future, the integration of organoids with cutting-edge technologies such as synthetic biology, AI, and gene editing heralds a transformative phase in biomedical innovation. This synergy holds promise for unlocking new therapeutic pathways, advancing drug development, and providing profound new insights into complex diseases. The potential of organoids to serve as platforms for rapid drug testing and functional experimentation, coupled with their compatibility with gene-editing techniques, signifies a quantum leap in medical research. In summary, organoid technology stands at the forefront of a revolution in the biomedical field. Its impact on human society will be profound, and its future applications appear boundless. As researchers and clinicians continue to harness the power of this technology, we anticipate a future replete with groundbreaking discoveries and innovative treatments that will ultimately reshape our approach to understanding and curing human diseases.

Organoid technology, the use of three-dimensional cultures derived from stem cells that closely mimic the architecture and functionality of native organs, represents a seminal advance in biomedical science, offering a revolutionary perspective on human physiology and pathology. 1 Organoid technology has profoundly impacted various fields, notably oncology and regenerative medicine, demonstrating unparalleled adaptability and precision. 2 Organoids, essentially miniature versions of organs, offer a highly physiologically relevant model for understanding human biology. This relevance is particularly critical in drug development, an area in which traditional models often fall short. 3 Compared to two-dimensional cell cultures or animal models, organoids provide a more accurate representation of human tissues, enabling more reliable and efficient drug screening and functional validation. 4 This feature is particularly valuable in the context of cancer research, where organoids can mimic the tumor microenvironment, providing insights into tumor-immune interactions and host-pathogen dynamics. 5 The clinical fidelity of organoids is higher than that of conventional models, as they can replicate the complex biological processes of human organs in vitro . This attribute enables the rapid functional testing of drugs, increasing the efficiency of the pathway from discovery to clinical application. 6 Additionally, organoids present a novel platform for in vitro gene-editing therapies. By leveraging CRISPR-Cas9 and other gene-editing tools, researchers can use organoids to model genetic diseases and test therapeutic strategies, significantly advancing the field of personalized medicine. 7 , 8 Our research findings emphasize the transformative potential of organoids in biomedical sciences. In the future, we expect organoids to play a pivotal role in advancing our understanding of human biology, evolving from cellular self-organization to integration with technologies such as synthetic biology and artificial intelligence (AI), offering substantial potential to revolutionize fields such as drug discovery, disease modeling, and personalized medicine. Progress in organoid technology continues to break barriers, offering insights once deemed unattainable. Accordingly, this review not only explores the current landscape of organoid technology but also offers a forward-looking perspective on the profound anticipated impact of organoids on future biomedical research.

Recent technological advances are further empowering organoid systems and enabling the derivation of new biological insights. New tools in single-cell omics, imaging, genetics, and AI are being integrated with organoid platforms. Single-cell RNA sequencing (scRNA-seq) and other omics tools enable the high-resolution dissection of organoid composition and function ( Figure 8 ). 222 Advanced imaging techniques allow the dynamic visualization of organoid morphogenesis and cell-cell interactions. 223 AI and machine learning provide computational power to extract complex patterns and build predictive models from multidimensional organoid data. 224 The integration of organoids with these emerging technologies has created a new generation of “organs-on-chips” that recapitulate tissue- and organ-level physiology in unprecedented detail. 225 , 226 Figure 8 Tech-infused organoids: The cutting-edge advancement in biomedicine Organoids are now being enhanced with cutting-edge technical advancements. The integration of several disciplines in joint efforts has led to the development of organoids that are equipped with cutting-edge technology. This advancement holds the potential to bring about a significant transformation in the field of biomedicine. Tech-infused organoids: The cutting-edge advancement in biomedicine Organoids are now being enhanced with cutting-edge technical advancements. The integration of several disciplines in joint efforts has led to the development of organoids that are equipped with cutting-edge technology. This advancement holds the potential to bring about a significant transformation in the field of biomedicine. Genetic engineering in organoids is a rapidly advancing field that combines organ-like complexity with molecular precision. Methods such as viral (retroviral, lentiviral, adenoviral) and nonviral (electroporation, lipofection) delivery systems enable gene manipulation in organoids, 227 and CRISPR-Cas9 is revolutionizing precise gene editing. 228 RNA interference is also used to modulate gene expression. 227 Organoids are vital in high-throughput screening for assessing cell viability and metabolic activity. Enzyme-linked immunosorbent assays quantify biomarkers, while image-based profiling techniques such as epifluorescence and confocal microscopy are used to analyze organoid morphology and functional changes after treatment. 163 , 229 High-content analysis dissects developmental processes and disease progression at the cellular level, integrating omics data and multidimensional imaging for a comprehensive view of organoid responses to drug treatments. z-stack imaging provides a complete cross-sectional view of three-dimensional organoids, increasing the accuracy of drug efficacy and toxicity assessments. 229 Computational methods model cellular interactions and biomechanical properties in organoids, facilitating the optimization of culture conditions and the prediction of drug-treatment outcomes. 230 Human liver organoids derived from PSCs have been successfully used to model drug-induced liver injury, aiding in antifibrosis drug screening and the identification of compounds with significant antifibrotic effects, such as SD208 and imatinib. 230 The integration of single-cell omics and organoid technologies has significantly advanced our understanding of organ development, disease mechanisms, and potential therapeutic approaches. This synergy has been especially effective in the field of neurobiology and brain organoid research. Fleck et al. investigated the gene-regulatory network underlying human cerebral organoid development and used multiomics single-cell data to develop an algorithm called Pando to infer the gene-regulatory network, thereby revealing the involvement of different transcription factors at various stages of cerebral organoid development. Their research emphasizes the conservation of developmental programs across species and the predictive capability of multiregion human cerebral organoids as model systems. 231 Vento-Tormo emphasized the revolutionary upgrade that organoid technology, 232 combined with single-cell profiling, has brought to the field of human development. This combination allows a deeper understanding of cell-cell communication and cellular heterogeneity, as evidenced by the work of Camp et al. in studying human liver organoids using scRNA-seq. 232 Yin et al. further elaborated on the strengths of combining scRNA-seq and organoid technologies, highlighting applications such as the discovery of novel cell types and gene markers, the recapitulation of cellular heterogeneity, the delineation of cell-differentiation pathways, and the modeling of diseases. 233 Single-cell omics technologies also enable the high-resolution dissection of cellular heterogeneity within organoid systems. scRNA-seq has been transformative in decoding organoid composition and identifying novel cell states. For example, scRNA-seq was used to map the cell lineages and transitional states in intestinal organoids, reconstructing the differentiation trajectories from stem cells to all intestinal epithelial cell types. 222 In patient-derived organoids, a recent study adopted an integrative approach combining scRNA-seq, epigenomic single-nucleotide polymorphism (SNP)-to-gene mapping, and genome-wide association study summary statistics to deduce the cellular subtypes and molecular pathways by which genetic variants affect disease phenotypes. 234 In summary, the integration of single-cell omics with organoid technologies provides a robust platform for modeling organ development and diseases. This combination has proven effective in revealing the cellular and molecular intricacies of organogenesis and disease pathogenesis. 235 These advancements not only deepen our understanding of human biology but also pave the way for the development of novel therapeutic strategies. The field of single-cell omics faces challenges in data interpretation due to the complexity of merging high-dimensional single-cell data with spatial information. The integration of AI and deep-learning algorithms can help streamline this process, enabling the identification of intricate patterns and interactions that are not easily discernible through traditional methods. Combining organoids with AI and deep learning enables innovative approaches to analyzing complex biological datasets and constructing predictive models. Organoids produce extensive multidimensional data, including cellular imaging, omics profiles, phenotypic readouts, and time series, and AI offers the computational power to extract meaningful patterns from this data overload. 236 One major application is automated image analysis using convolutional neural networks, which can segment and classify cells in organoid imaging data orders of magnitude faster than human annotation. 237 AI also enables phenotypic profiling from images. A deep-learning model was trained to predict the emergence of retinal tissue morphology and cell types in optic cup organoids over time. 238 In the absence of reliable markers for assessing organoid vitality, researchers recently devised an innovative approach using AI algorithms to analyze a spectrum of phenotypic parameters. Remarkably, the AI-driven method can ascertain organoid aging solely through bright-field imaging. This approach eliminates the cumbersome steps of preparing single-cell suspensions and performing senescence-associated β-galactosidase staining, thereby streamlining quality control processes and making biobank operations more efficient. 224 Beyond merely visualizing images, AI can concurrently reconstruct multiple parameters from multiomics data in organoids. These data are then integrated with complementary datasets, such as functional metrics and patient records, all derived from the same experimental conditions. The resulting repository is a large multimodal database. AI algorithms sift through this complex data to identify patterns and correlations, elucidating connections within diverse multiparametric information that may be challenging for human analysts to interpret. Overall, the integration of organoids with AI and deep learning constitutes an exciting new frontier. The integration of organoids with microfluidics and sensors has enabled the development of “organs-on-chips” that recapitulate key aspects of human tissue physiology in vitro . These biomimetic systems incorporate organoids into microengineered devices with tissue-tissue interfaces, fluid flow, and real-time monitoring to model organ function and disease. 225 One example is the gut-on-a-chip, which combines intestinal organoids with a microfluidic device lined by endothelial and immune cells to model the intestinal epithelium. This system faithfully reproduces the microenvironment of the living intestine, including peristalsis-like motions and cyclic strain from breathing. Using this model, researchers showed that cyclic strain promotes intestinal stem cell differentiation and epithelial permeability. Thus, the gut-on-a-chip provides a powerful platform to study intestinal absorption, inflammation, gut-microbe interactions, and more. 239 , 240 , 241 Another study by Campisi et al. presents a three-dimensional self-organized microvascular model of the blood-brain barrier using human iPSC-derived endothelial cells, brain pericytes, and astrocytes in a fibrin gel. This microfluidic system replicates in vivo neurovascular organization, exhibiting lower permeability than conventional in vitro models, and thus provides a physiologically relevant and robust platform for drug discovery and the prediction of neurotherapeutic transport efficacy. 242 Hajal et al. developed a human blood-brain barrier model within microfluidic devices by incorporating endothelial cells, brain pericytes, and astrocytes in a three-dimensional gel matrix. This model forms three-dimensional vessel architectures that resemble the natural blood-brain barrier, with gene-expression profiles and permeability values comparable to those observed in vivo . 243 Accordingly, this blood-brain barrier model facilitates the quantitative assessment of molecular permeabilities and is suitable for use in both academic and industrial research to predict transport across the blood-brain barrier. These studies exemplify significant advances in organ-on-a-chip technology, specifically in modeling the blood-brain barrier. 244 This progress underscores the potential of organ-on-a-chip technology to revolutionize biomedical research and therapeutic development. Another exciting emerging application of organoids is the modeling of neural networks for biocomputation. Like their in vivo counterparts, the organized neuronal networks generated within brain organoids can perform specialized information processing. 245 Researchers now plan to develop brain organoids that can serve as “biocomputers” for tasks such as pattern recognition and classification. 246 In one pioneering study, a proof of concept of using brain organoids for biosensing and biocomputation was demonstrated. A recent publication in the journal Neuron revealed groundbreaking research conducted by Brett Kagan and his team at Cortical Labs, demonstrating that organoid neural networks cultured in vitro can learn to play the classic arcade game Pong. 247 These so-called mini-brains, composed of interconnected neural cells, exhibit the ability to perceive and interact with their surrounding environment. Utilizing a computer interface, the neural networks were trained to detect the position of the game’s digital sphere and manipulate a virtual paddle accordingly. Remarkably, the assemblage of brain cells acquired this skill within 5 min. Biocomputing with brain organoids could potentially be faster, more efficient, and more powerful than silicon-based computing and AI, requiring only a fraction of the energy. 248 This technology could enable brain-inspired AI and powerful models for studying neural information processing. Looking forward, the integration of organoids with new technologies has immense potential to transform biomedicine and human health. Such advances could power a new generation of preclinical disease models for drug screening, enable regenerative therapies using laboratory-grown tissues, and provide platforms for precision medicine. Realizing this potential will require collaborative multidisciplinary efforts at the intersection of stem cell biology, bioengineering, and data science. An exciting future lies ahead as organoids and emerging technologies combine to elucidate human biology in unprecedented detail.