Human spinal cord organoids(hSCOs)offer a promising platform to study neurotrauma by addressing many limitations of traditional research models.These organoids provide access to human-specific physiological and geneti...Human spinal cord organoids(hSCOs)offer a promising platform to study neurotrauma by addressing many limitations of traditional research models.These organoids provide access to human-specific physiological and genetic mechanisms and can be derived from an individual's somatic cells(e.g.,blood or skin).This enables patient-specific paradigms for precision neurotrauma research,pa rticula rly relevant to the over 300,000 people in the United States living with chronic effects of spinal cord injury(SCI).展开更多
Human cardiac organoids have revolutionized the study of cardiac development,disease modeling, drug discovery, and regenerative therapies. This review systematically discusses strategies and progress in the constructi...Human cardiac organoids have revolutionized the study of cardiac development,disease modeling, drug discovery, and regenerative therapies. This review systematically discusses strategies and progress in the construction of cardiac organoids, categorizing them into three main types:cardiac spheroids, self-organizing/assembloid organoids, and organoid-on-a-chip systems. This review uniquely integrates the advances in vascularization, organ-on-chip design, and environmental cardiotoxicity modeling within cardiac organoid platforms, offering a critical synthesis that is absent in the literature. In the context of escalating environmental threats to cardiovascular health, there is an urgent need for physiologically relevant models to accurately identify cardiac toxicants and elucidate their underlying mechanisms of action. This review highlights advances in cardiac organoid applications for disease modeling-including congenital heart defects and acquired cardiovascular diseases-drug development, toxicity screening, and the study of environmentally induced cardiovascular pathogenesis. In addition, it critically examines ongoing challenges and underscores opportunities brought by bioengineering approaches. Finally, we propose future directions for developing standardized cardiac organoid platforms with clinical predictability, aiming to expand the utility of this technology across broader research applications.展开更多
Organoids possess immense potential for unraveling the intricate functions of human tissues and facilitating preclinical disease treatment.Their applications span from high-throughput drug screening to the modeling of...Organoids possess immense potential for unraveling the intricate functions of human tissues and facilitating preclinical disease treatment.Their applications span from high-throughput drug screening to the modeling of complex diseases,with some even achieving clinical translation.Changes in the overall size,shape,boundary,and other morphological features of organoids provide a noninvasive method for assessing organoid drug sensitivity.However,the precise segmentation of organoids in bright-field microscopy images is made difficult by the complexity of the organoid morphology and interference,including overlapping organoids,bubbles,dust particles,and cell fragments.This paper introduces the precision organoid segmentation technique(POST),which is a deep-learning algorithm for segmenting challenging organoids under simple bright-field imaging conditions.Unlike existing methods,POST accurately segments each organoid and eliminates various artifacts encountered during organoid culturing and imaging.Furthermore,it is sensitive to and aligns with measurements of organoid activity in drug sensitivity experiments.POST is expected to be a valuable tool for drug screening using organoids owing to its capability of automatically and rapidly eliminating interfering substances and thereby streamlining the organoid analysis and drug screening process.展开更多
Background:Cartilage repair remains a considerable challenge in regenerative medicine.Despite extensive research on biomaterials for cartilage repair in recent years,issues such as prolonged repair cycles and suboptim...Background:Cartilage repair remains a considerable challenge in regenerative medicine.Despite extensive research on biomaterials for cartilage repair in recent years,issues such as prolonged repair cycles and suboptimal outcomes persist.Organoids,miniature three-dimensional(3D)tissue structures derived from the directed differentiation of stem or progenitor cells,mimic the structure and function of natural organs.Therefore,the construction of cartilage organoids(COs)holds great promise as a novel strategy for cartilage repair.Methods:This study employed a digital light processing system to perform 3D bioprinting of a DNA-silk fibroin(DNA-SF)hydrogel sustained-release system(DSRGT)with bone-marrow mesenchymal stem cells(BMSCs)to construct millimeter-scale CO.COs at different developmental stages were characterized,and the COs with the best cartilage phenotype were selected for in vivo cartilage repair in a rat articular cartilage defect model.Results:This study developed a DSRGT by covalently grafting glucosamine(which promotes cartilage matrix synthesis)and TD-198946(which promotes chondrogenic differentiation)onto a hydrogel using acrylic acid-polyethylene glycolN-hydroxysuccinimide(AC-PEG-NHS).In vitro,4-week COs exhibited higher SRY-box transcription factor 9(SOX9),typeⅡcollagen(ColⅡ),and aggrecan(ACAN)expression and lower typeⅠcollagen(ColⅠ)and typeⅩcollagen(ColⅩ)expression,indicating that 4 weeks is the optimal culture duration for hyaline cartilage development.In vivo,the mitogen-activated protein kinase(MAPK)signaling pathway was upregulated in 4-week COs,enabling cartilage repair within 8 weeks.Transcriptomic analysis revealed that cartilage regenerated with 4-week COs presented gene expression profiles resembling those of healthy cartilage.Conclusion:This study employs DSRGT to construct COs,providing an innovative strategy for the regeneration of cartilage defects.展开更多
Background:Hepatocellular carcinoma(HCC)is a highly lethal malignancy driven by both intrinsic oncogenic pathways and immune microenvironmental regulation.Emerging evidence suggests that DNASE1L3 may influence tumor b...Background:Hepatocellular carcinoma(HCC)is a highly lethal malignancy driven by both intrinsic oncogenic pathways and immune microenvironmental regulation.Emerging evidence suggests that DNASE1L3 may influence tumor biology and immune responses;however,its specific roles in HCC progression and macrophage-mediated regulation remain unclear.This study aimed to elucidate the biological functions of DNASE1L3 in HCC and to determine how it modulates tumor behavior and immune interactions.Methods:Bioinformatics analyses of the GSE41804 and Cancer Genome Atlas-Liver Hepatocellular Carcinoma(TCGA-LIHC)datasets were used to identify hub genes.Functional assays assessed the impact of DNASE1L3 on HCC cell proliferation,migration,invasion,and cell cycle progression.The effects of DNASE1L3 on macrophage polarization and the Wnt/β-catenin signaling pathway were examined using a co-culture system.An HCC organoid model was established to further validate its regulatory function.Results:Eight prognostic signature genes were identified,with deoxyribonuclease I-like 3(DNase I-like 3)selected as the hub gene.DNASE1L3 overexpression suppressed HCC cell growth,inhibited migration and invasion,induced G1 arrest,and modulated epithelial-mesenchymal transition(EMT)markers.DNASE1L3 knockdown promoted M2-like macrophage polarization.Mechanistically,DNASE1L3 interacted withβ-catenin to enhance its ubiquitination and degradation,thereby inhibiting Wnt/β-catenin signaling and reducing PD-L1 expression.DNASE1L3 overexpression similarly restricted organoid growth and suppressed pathway activity.Conclusion:DNASE1L3 acts as a negative regulator of HCC progression by targeting the Wnt/β-catenin pathway and reducing PD-L1 expression,thereby influencing both tumor cell behavior and macrophage-mediated immune responses.展开更多
The brain is the most complex human organ,and commonly used models,such as two-dimensional-cell cultures and animal brains,often lack the sophistication needed to accurately use in research.In this context,human cereb...The brain is the most complex human organ,and commonly used models,such as two-dimensional-cell cultures and animal brains,often lack the sophistication needed to accurately use in research.In this context,human cerebral organoids have emerged as valuable tools offering a more complex,versatile,and human-relevant system than traditional animal models,which are often unable to replicate the intricate architecture and functionality of the human brain.Since human cerebral organoids are a state-of-the-art model for the study of neurodevelopment and different pathologies affecting the brain,this field is currently under constant development,and work in this area is abundant.In this review,we give a complete overview of human cerebral organoids technology,starting from the different types of protocols that exist to generate different human cerebral organoids.We continue with the use of brain organoids for the study of brain pathologies,highlighting neurodevelopmental,psychiatric,neurodegenerative,brain tumor,and infectious diseases.Because of the potential value of human cerebral organoids,we describe their use in transplantation,drug screening,and toxicology assays.We also discuss the technologies available to study cell diversity and physiological characteristics of organoids.Finally,we summarize the limitations that currently exist in the field,such as the development of vasculature and microglia,and highlight some of the novel approaches being pursued through bioengineering.展开更多
Brain organoids encompass a large collection of in vitro stem cell–derived 3D culture systems that aim to recapitulate multiple aspects of in vivo brain development and function.First,this review provides a brief int...Brain organoids encompass a large collection of in vitro stem cell–derived 3D culture systems that aim to recapitulate multiple aspects of in vivo brain development and function.First,this review provides a brief introduction to the current state-of-the-art for neuroectoderm brain organoid development,emphasizing their biggest advantages in comparison with classical two-dimensional cell cultures and animal models.However,despite their usefulness for developmental studies,a major limitation for most brain organoid models is the absence of contributing cell types from endodermal and mesodermal origin.As such,current research is highly investing towards the incorporation of a functional vasculature and the microglial immune component.In this review,we will specifically focus on the development of immune-competent brain organoids.By summarizing the different approaches applied to incorporate microglia,it is highlighted that immune-competent brain organoids are not only important for studying neuronal network formation,but also offer a clear future as a new tool to study inflammatory responses in vitro in 3D in a brainlike environment.Therefore,our main focus here is to provide a comprehensive overview of assays to measure microglial phenotype and function within brain organoids,with an outlook on how these findings could better understand neuronal network development or restoration,as well as the influence of physical stress on microglia-containing brain organoids.Finally,we would like to stress that even though the development of immune-competent brain organoids has largely evolved over the past decade,their full potential as a pre-clinical tool to study novel therapeutic approaches to halt or reduce inflammation-mediated neurodegeneration still needs to be explored and validated.展开更多
An organoid is a three-dimensional(3D)cell culture model that can reproduce the distinct structure and inherent functionality of certain organs.Nevertheless,a major limitation of organoids is the absence of a complex ...An organoid is a three-dimensional(3D)cell culture model that can reproduce the distinct structure and inherent functionality of certain organs.Nevertheless,a major limitation of organoids is the absence of a complex vascular network,thus restricting the supply of oxygen and essential nutrients.Coupled with their inherent size constraints and metabolite accumulation,it is challenging for organoids to replicate the natural intricacies of organs,thereby limiting their applicability.To overcome the challenges associated with this technology,we developed a culture platform to cultivate tumors or organ-derived organoids up to the centimeter scale.Initially,a customized organoid-on-a-chip including a microvascular network at the micron scale was designed using 3D printing.Further,by integrating an infusion device,the chip ensures an adequate supply of nutrients and fluid immersion while mimicking blood flow dynamics.Our method overcomes the issue of the limited size of organoids due to insufficient nutrient access,making it possible to produce large-scale tumor and normal tissue models in vitro,while providing insights into drug efficacy and toxicology evaluation as well as standardized organoid production.展开更多
The structure of intestinal tissue is complex.In vitro simulation of intestinal structure and function is important for studying intestinal development and diseases.Recently,organoids have been successfully constructe...The structure of intestinal tissue is complex.In vitro simulation of intestinal structure and function is important for studying intestinal development and diseases.Recently,organoids have been successfully constructed and they have come to play an important role in biomedical research.Organoids are miniaturized three-dimensional(3D)organs,derived from stem cells,which mimic the structure,cell types,and physiological functions of an organ,making them robust models for biomedical research.Intestinal organoids are 3D micro-organs derived from intestinal stem cells or pluripotent stem cells that can successfully simulate the complex structure and function of the intestine,thereby providing a valuable platform for intestinal development and disease research.In this article,we review the latest progress in the construction and application of intestinal organoids.展开更多
Current organoid-generation strategies rely predominantly on intricate in vitro manipulations of dissociated stem cells,including isolation,expansion,and genetic modification.However,these approaches present significa...Current organoid-generation strategies rely predominantly on intricate in vitro manipulations of dissociated stem cells,including isolation,expansion,and genetic modification.However,these approaches present significant challenges in terms of safety and scalability for clinical applications.An alternative strategy involves the direct generation of organoids from readily available tissues.Herein,we report the generation of functional organoids representing all three germ layers from human adult adipose tissue without single-cell processing steps.Specifically,by employing a specialized suspension culture system,we have developed reaggregated microfat(RMF)tissues,which differentiated into mesodermal bone marrow organoids capable of reconstituting human normal hematopoiesis in immunodeficient mice,endodermal insulin-producing organoids that reversed hyperglycemia in streptozotocin(STZ)-induced diabetic mice,and ectodermal nervous-like tissues resembling neurons and neuroglial cells.These findings therefore highlight the potential of human adipose tissue as a safe,scalable,and clinically viable source for organoid-based regenerative therapies.展开更多
Hypoxic injury(HI)in the prenatal period often causes neonatal neurological disabilities.Due to the difficulty in obtaining clinical samples,the molecular and cellular mechanisms remain unclear.Here we use vascularize...Hypoxic injury(HI)in the prenatal period often causes neonatal neurological disabilities.Due to the difficulty in obtaining clinical samples,the molecular and cellular mechanisms remain unclear.Here we use vascularized cerebral organoids to investigate the hypoxic injury phenotype and explore the intercellular interactions between vascular and neural tissues under hypoxic conditions.Our results indicate that fused vascularized cerebral organoids exhibit broader hypoxic responses and larger decreases in panels of neural development-related genes when exposed to low oxygen levels compared to single cerebral organoids.Interestingly,vessels also exhibit neural protective effects on T-box brain protein 2+intermediate progenitors(IPs),which are markedly lost in HI cerebral organoids.Furthermore,we identify the role of bone morphogenic protein signaling in protecting IPs.Thus,this study has established an in vitro organoid system that can be used to study the contribution of vessels to brain injury under hypoxic conditions and provides a strategy for the identification of intervention targets.展开更多
Brain organoid-on-chip platforms have emerged as groundbreaking tools in neural disease modeling and drug discovery,offering a unique and highly accurate simulation of human organ physiology and func-tion compared wit...Brain organoid-on-chip platforms have emerged as groundbreaking tools in neural disease modeling and drug discovery,offering a unique and highly accurate simulation of human organ physiology and func-tion compared with traditional cell culture systems.This technology is a harmonious fusion of organ-on-a-chip and organoid culture technologies,leveraging their strengths to provide the most realistic in vitro replication of the in vivo environment,both physically and biologically.As both technologies continue to advance rapidly,this platform is highly promising in vitro platform for disease modeling.In this review,we summarize the historical developments,recent advancements,limitations,and future prospects of brain organoid-on-chip technology,aiming to illuminate the transformative potential of this platform in advancing our understanding and treatment of neural diseases.展开更多
Human brain development is a complex process,and animal models often have significant limitations.To address this,researchers have developed pluripotent stem cell-derived three-dimensional structures,known as brain-li...Human brain development is a complex process,and animal models often have significant limitations.To address this,researchers have developed pluripotent stem cell-derived three-dimensional structures,known as brain-like organoids,to more accurately model early human brain development and disease.To enable more consistent and intuitive reproduction of early brain development,in this study,we incorporated forebrain organoid culture technology into the traditional unguided method of brain organoid culture.This involved embedding organoids in matrigel for only 7 days during the rapid expansion phase of the neural epithelium and then removing them from the matrigel for further cultivation,resulting in a new type of human brain organoid system.This cerebral organoid system replicated the temporospatial characteristics of early human brain development,including neuroepithelium derivation,neural progenitor cell production and maintenance,neuron differentiation and migration,and cortical layer patterning and formation,providing more consistent and reproducible organoids for developmental modeling and toxicology testing.As a proof of concept,we applied the heavy metal cadmium to this newly improved organoid system to test whether it could be used to evaluate the neurotoxicity of environmental toxins.Brain organoids exposed to cadmium for 7 or 14 days manifested severe damage and abnormalities in their neurodevelopmental patterns,including bursts of cortical cell death and premature differentiation.Cadmium exposure caused progressive depletion of neural progenitor cells and loss of organoid integrity,accompanied by compensatory cell proliferation at ectopic locations.The convenience,flexibility,and controllability of this newly developed organoid platform make it a powerful and affordable alternative to animal models for use in neurodevelopmental,neurological,and neurotoxicological studies.展开更多
Organoids are three-dimensional stem cell-derived models that offer a more physiologically relevant representation of tumor biology compared to traditional two-dimensional cell cultures or animal models.Organoids pres...Organoids are three-dimensional stem cell-derived models that offer a more physiologically relevant representation of tumor biology compared to traditional two-dimensional cell cultures or animal models.Organoids preserve the complex tissue architecture and cellular diversity of human cancers,enabling more accurate predictions of tumor growth,metastasis,and drug responses.Integration with microfluidic platforms,such as organ-on-a-chip systems,further enhances the ability to model tumor-environment interactions in real-time.Organoids facilitate in-depth exploration of tumor heterogeneity,molecular mechanisms,and the development of personalized treatment strategies when coupled with multi-omics technologies.Organoids provide a platform for investigating tumor-immune cell interactions,which aid in the design and testing of immune-based therapies and vaccines.Taken together,these features position organoids as a transformative tool in advancing cancer research and precision medicine.展开更多
Coronavirus disease 2019(COVID-19),caused by severe acute respiratory syndrome coronavirus 2(SARS-CoV-2),not only affects the lungs but also damages various non-pulmonary organs,resulting in tissue injury and potentia...Coronavirus disease 2019(COVID-19),caused by severe acute respiratory syndrome coronavirus 2(SARS-CoV-2),not only affects the lungs but also damages various non-pulmonary organs,resulting in tissue injury and potential long-term sequelae in infected individuals.COVID-19 is likely to persist as a public health concern,given the frequent emergence of new mutations and viral strains.Multiple clinical lines of evidence indicate the efficacy of traditional Chinese medicine(TCM)in the prevention and treatment of COVID-19.However,the exact mechanism underlying these effects remains unclear.In this perspective review,we summarize the utility of in vitro three-dimensional(3D)cultured organoid models and organ-on-a-chip(OoC)technology for studying COVID-19 pathogenesis,viral tropism,and infectious mechanisms across different tissues.We highlight the successful application of these platforms in aiding drug development and discuss their advantages and limitations.We also review how such organotypic models can be employed to study TCMs.Finally,we discuss the opportunities for integrated microphysiological multi-tissue models to rapidly discover active components and potential targets in the context of COVID-19.The utilization of these emerging technologies could accelerate drug discovery and the modernization of TCM.展开更多
BACKGROUND Hepatic organoid-based modelling,through the elucidation of a range of in vivo biological processes and the recreation of the intricate liver microenvironment,is yielding groundbreaking insights into the pa...BACKGROUND Hepatic organoid-based modelling,through the elucidation of a range of in vivo biological processes and the recreation of the intricate liver microenvironment,is yielding groundbreaking insights into the pathophysiology and personalized medicine approaches for liver diseases.AIM This study was designed to analyse the global scientific output of hepatic organoid research and assess current achievements and future trends through bibliometric analysis.METHODS Articles were retrieved from the Web of Science Core Collection,and CiteSpace 6.3.R1 was employed to analyse the literature,including outputs,journals,and countries,among others.RESULTS Between 2010 and 2024,a total of 991 articles pertaining to hepatic organoid research were published.The journal Hepatology published the greatest number of papers,and journals with an impact factor greater than 10 constituted 60%of the top 10 journals.The United States and Utrecht University were identified as the most prolific country and institution,respectively.Clevers H emerged as the most prolific author,whereas Huch M had the highest number of cocitations,suggesting that both are ideal candidates for academic collaboration.Research on hepatic organoids has exhibited a progressive shift in focus,evolving from initial investigations into model building,differentiation research in stem cells,bile ducts,and progenitor cells,to a broader spectrum encompassing lipid metabolism,single-cell RNA sequencing,and therapeutic applications.The phrases exhibiting citation bursts from 2022 to 2024 include“drug resistance”,“disease model”,and“patient-derived tumor organoids”.CONCLUSION Research on hepatic organoids has increased over the past decade and is expected to continue to grow.Key research areas include applications for liver diseases and drug development.Future trends likely to gain focus include patient-derived tumour organoids,disease modelling,and personalized medicine.展开更多
Liposarcoma is one of the most common soft tissue sarcomas,however,its occurrence rate is still rare compared to other cancers.Due to its rarity,in vitro experiments are an essential approach to elucidate liposarcoma ...Liposarcoma is one of the most common soft tissue sarcomas,however,its occurrence rate is still rare compared to other cancers.Due to its rarity,in vitro experiments are an essential approach to elucidate liposarcoma pathobiology.Conventional cell culture-based research(2D cell culture)is still playing a pivotal role,while several shortcomings have been recently under discussion.In vivo,mouse models are usually adopted for pre-clinical analyses with expectations to overcome the issues of 2D cell culture.However,they do not fully recapitulate human dedifferentiated liposarcoma(DDLPS)characteristics.Therefore,three-dimensional(3D)culture systems have been the recent research focus in the cell biology field with the expectation to overcome at the same time the disadvantages of 2D cell culture and in vivo animal models and fill in the gap between them.Given the liposarcoma rarity,we believe that 3D cell culture techniques,including 3D cell cultures/co-cultures,and Patient-Derived tumor Organoids(PDOs),represent a promising approach to facilitate liposarcoma investigation and elucidate its molecular mechanisms and effective therapy development.In this review,we first provide a general overview of 3D cell cultures compared to 2D cell cultures.We then focus on one of the recent 3D cell culture applications,Patient-Derived Organoids(PDOs),summarizing and discussing several PDO methodologies.Finally,we discuss the current and future applications of PDOs to sarcoma,particularly in the field of liposarcoma.展开更多
BACKGROUND Glioblastoma multiforme(GBM)is the most aggressive and prevalent primary malignant brain tumor in adults,marked by poor prognosis and high invasiveness.Traditional GBM invasion assays,such as those involvin...BACKGROUND Glioblastoma multiforme(GBM)is the most aggressive and prevalent primary malignant brain tumor in adults,marked by poor prognosis and high invasiveness.Traditional GBM invasion assays,such as those involving mouse brain xenografts,are often time-consuming and limited in efficiency.In this context,stem cell-derived neural organoids(NOs)have emerged as advanced,threedimensional,human-relevant platforms that mimic the cellular architecture and microenvironment of the human brain.These models provide novel opportunities to investigate glioblastoma stem cell invasion,a critical driver of tumor progression and therapeutic resistance.AIM To evaluate studies using stem cell-derived NOs to model glioblastoma migration/invasion,focusing on methodologies,applications and therapeutic implications.METHODS We conducted a systematic review following PRISMA guidelines,searching PubMed and Scopus for studies published between March 2019 and March 2025 that investigated NOs in the context of glioblastoma invasion/migration.After screening 377 articles based on predefined inclusion and exclusion criteria,10 original research articles were selected for analysis.Extracted data were categorized into four analytical domains:(1)Tumor model formation;(2)NO characteristics;(3)NO differentiation protocols;and(4)Invasion/migration assessment methodologies.RESULTS The included studies exhibit significant methodological heterogeneity GBM model development,particularly regarding model type,cell source and culture conditions.Most studies(70%)used suspension cell models,while 30%employed spheroids,with most research focusing on patient-derived glioblastoma stem cells.NOs were predominantly generated from human induced pluripotent stem cells using both guided and unguided differentiation protocols.Confocal fluorescence microscopy was the primary method used for assessing invasion,revealing invasion depths of up to 300μm.Organoid maturity and co-culture duration influenced results,while key factors for model optimization included tumor cell density,organoid age and extracellular matrix composition.Some studies also tested therapeutic strategies such as Zika virus and microRNA modulation.Collectively,findings support the utility of NOs as effective tools for studying GBM behavior and therapeutic responses in a humanized three-dimensional context.CONCLUSION Human NOs represent promising platforms for modeling glioblastoma invasion in a humanized three-dimensional environment.However,a limited number of studies and methodological heterogeneity hinder reproducibility.Protocol standardization is essential to enhance the translational application of these models.展开更多
The neurotransmitter 5-hydroxytryptamine(5-HT),primarily produced by intestinal enterochromaffin(EC)cells,relies on tryptophan hydroxylase 1(TPH1)for synthesis.Research suggested Bifidobacterium breve CCFM1025’s pote...The neurotransmitter 5-hydroxytryptamine(5-HT),primarily produced by intestinal enterochromaffin(EC)cells,relies on tryptophan hydroxylase 1(TPH1)for synthesis.Research suggested Bifidobacterium breve CCFM1025’s potential in regulating Tph1 gene expression,maintaining 5-HT levels in stressed mice,but its precise mechanisms were unclear.This study used metabolomic techniques to assess probiotic fermentation products,revealing acetate as the crucial element in Bb-CCFM1025’s regulation of gut 5-HT synthesis.Further exploration correlated acetate with Tph1 transcription in intestinal organoids.Transcriptomic methods and quantitative reverse transcription polymerase chain reaction validation demonstrated how acetate facilitated 5-HT synthesis and secretion.It unveiled that acetate orchestrates signaling pathways(phosphoinositide 3-kinase-protein kinase B(PI3K-AKT),phospholipase C-phosphorylated extracellular signal-regulated kinase(PLC-pERK),and PLC-1,4,5-trisphosphate(IP3)-Ca^(2+))within EC,enabling 5-HT production.These findings elucidate the biochemical mechanisms behind specific probiotics’effects,aiding in the targeted selection of similar beneficial strains.This study offers theoretical support for choosing probiotics with analogous functionalities based on their physiological impacts.展开更多
Organoids are derived from self-organizing stem cells and form three-dimensional structures that are structurally and functionally similar to in vivo tissues.With the ability to replicate the in vivo microenvironment ...Organoids are derived from self-organizing stem cells and form three-dimensional structures that are structurally and functionally similar to in vivo tissues.With the ability to replicate the in vivo microenvironment and maintain genetic stability,organoids have become a powerful tool for elucidating developmental mechanisms,accurately modeling disease processes,and efficiently screening drug candidates,and have also demonstrated significant value in the field of traditional Chinese medicine(TCM)-including applications in screening active components of TCM,studying TCM pharmacodynamic mechanisms,evaluating TCM safety,and verifying the effects of traditional non-pharmacological therapies such as acupuncture and yoga.Organoids can be cultured using air-liquid interface systems,bioreactors,and vascularization techniques.They are widely used in drug screening,disease modeling,precision medicine,and toxicity assessment.However,current limitations include high costs,difficulty in accurately replicating the microenvironment,and ethical concerns.In this review,we systematically retrieve,synthesize,and analyze relevant literature to elucidate the culture methods of organoid technology,its diverse applications across various fields,and the challenges it faces.In the future,integration with artificial intelligence may provide new insights and strategies for drug development and disease research and the modernization of TCM.展开更多
基金supported by the Belle Carnell Regenerative Neurorehabilitation Fundthe National Institutes of Health(R01NS113935 to CKF)。
文摘Human spinal cord organoids(hSCOs)offer a promising platform to study neurotrauma by addressing many limitations of traditional research models.These organoids provide access to human-specific physiological and genetic mechanisms and can be derived from an individual's somatic cells(e.g.,blood or skin).This enables patient-specific paradigms for precision neurotrauma research,pa rticula rly relevant to the over 300,000 people in the United States living with chronic effects of spinal cord injury(SCI).
基金supported by the Innovation Promotion Program of NHC and Shanghai Key Labs,SIBPT(grant number PT2025-01)。
文摘Human cardiac organoids have revolutionized the study of cardiac development,disease modeling, drug discovery, and regenerative therapies. This review systematically discusses strategies and progress in the construction of cardiac organoids, categorizing them into three main types:cardiac spheroids, self-organizing/assembloid organoids, and organoid-on-a-chip systems. This review uniquely integrates the advances in vascularization, organ-on-chip design, and environmental cardiotoxicity modeling within cardiac organoid platforms, offering a critical synthesis that is absent in the literature. In the context of escalating environmental threats to cardiovascular health, there is an urgent need for physiologically relevant models to accurately identify cardiac toxicants and elucidate their underlying mechanisms of action. This review highlights advances in cardiac organoid applications for disease modeling-including congenital heart defects and acquired cardiovascular diseases-drug development, toxicity screening, and the study of environmentally induced cardiovascular pathogenesis. In addition, it critically examines ongoing challenges and underscores opportunities brought by bioengineering approaches. Finally, we propose future directions for developing standardized cardiac organoid platforms with clinical predictability, aiming to expand the utility of this technology across broader research applications.
基金supported by the National Key R&D Program of China(No.2022YFC2504403)the National Natural Science Foundation of China(No.62172202)+1 种基金the Experiment Project of China Manned Space Program(No.HYZHXM01019)the Fundamental Research Funds for the Central Universities from Southeast University(No.3207032101C3)。
文摘Organoids possess immense potential for unraveling the intricate functions of human tissues and facilitating preclinical disease treatment.Their applications span from high-throughput drug screening to the modeling of complex diseases,with some even achieving clinical translation.Changes in the overall size,shape,boundary,and other morphological features of organoids provide a noninvasive method for assessing organoid drug sensitivity.However,the precise segmentation of organoids in bright-field microscopy images is made difficult by the complexity of the organoid morphology and interference,including overlapping organoids,bubbles,dust particles,and cell fragments.This paper introduces the precision organoid segmentation technique(POST),which is a deep-learning algorithm for segmenting challenging organoids under simple bright-field imaging conditions.Unlike existing methods,POST accurately segments each organoid and eliminates various artifacts encountered during organoid culturing and imaging.Furthermore,it is sensitive to and aligns with measurements of organoid activity in drug sensitivity experiments.POST is expected to be a valuable tool for drug screening using organoids owing to its capability of automatically and rapidly eliminating interfering substances and thereby streamlining the organoid analysis and drug screening process.
基金supported by the National Key Research and Development Program of China(2022YFB3804300)the National Natural Science Foundation of China(82230071,32471395,82427809,82472479)+2 种基金the Shanghai Science and Technology Innovation Action Plan(23141900600)the Research Physician Training Program of Shanghai Hospital Development Center(SHDC2023CRT013)the Shanghai Municipal Demonstration Project for Innovative Medical Device Applications(23SHS05700)。
文摘Background:Cartilage repair remains a considerable challenge in regenerative medicine.Despite extensive research on biomaterials for cartilage repair in recent years,issues such as prolonged repair cycles and suboptimal outcomes persist.Organoids,miniature three-dimensional(3D)tissue structures derived from the directed differentiation of stem or progenitor cells,mimic the structure and function of natural organs.Therefore,the construction of cartilage organoids(COs)holds great promise as a novel strategy for cartilage repair.Methods:This study employed a digital light processing system to perform 3D bioprinting of a DNA-silk fibroin(DNA-SF)hydrogel sustained-release system(DSRGT)with bone-marrow mesenchymal stem cells(BMSCs)to construct millimeter-scale CO.COs at different developmental stages were characterized,and the COs with the best cartilage phenotype were selected for in vivo cartilage repair in a rat articular cartilage defect model.Results:This study developed a DSRGT by covalently grafting glucosamine(which promotes cartilage matrix synthesis)and TD-198946(which promotes chondrogenic differentiation)onto a hydrogel using acrylic acid-polyethylene glycolN-hydroxysuccinimide(AC-PEG-NHS).In vitro,4-week COs exhibited higher SRY-box transcription factor 9(SOX9),typeⅡcollagen(ColⅡ),and aggrecan(ACAN)expression and lower typeⅠcollagen(ColⅠ)and typeⅩcollagen(ColⅩ)expression,indicating that 4 weeks is the optimal culture duration for hyaline cartilage development.In vivo,the mitogen-activated protein kinase(MAPK)signaling pathway was upregulated in 4-week COs,enabling cartilage repair within 8 weeks.Transcriptomic analysis revealed that cartilage regenerated with 4-week COs presented gene expression profiles resembling those of healthy cartilage.Conclusion:This study employs DSRGT to construct COs,providing an innovative strategy for the regeneration of cartilage defects.
基金funded by Shanghai Science and Technology Innovation Action Plan Project(22140901100)Shanghai Key Laboratory of Molecular Imaging(18DZ2260400)Shanghai University of Medicine and Health Science Seed Fund(SSF-24-21-01).
文摘Background:Hepatocellular carcinoma(HCC)is a highly lethal malignancy driven by both intrinsic oncogenic pathways and immune microenvironmental regulation.Emerging evidence suggests that DNASE1L3 may influence tumor biology and immune responses;however,its specific roles in HCC progression and macrophage-mediated regulation remain unclear.This study aimed to elucidate the biological functions of DNASE1L3 in HCC and to determine how it modulates tumor behavior and immune interactions.Methods:Bioinformatics analyses of the GSE41804 and Cancer Genome Atlas-Liver Hepatocellular Carcinoma(TCGA-LIHC)datasets were used to identify hub genes.Functional assays assessed the impact of DNASE1L3 on HCC cell proliferation,migration,invasion,and cell cycle progression.The effects of DNASE1L3 on macrophage polarization and the Wnt/β-catenin signaling pathway were examined using a co-culture system.An HCC organoid model was established to further validate its regulatory function.Results:Eight prognostic signature genes were identified,with deoxyribonuclease I-like 3(DNase I-like 3)selected as the hub gene.DNASE1L3 overexpression suppressed HCC cell growth,inhibited migration and invasion,induced G1 arrest,and modulated epithelial-mesenchymal transition(EMT)markers.DNASE1L3 knockdown promoted M2-like macrophage polarization.Mechanistically,DNASE1L3 interacted withβ-catenin to enhance its ubiquitination and degradation,thereby inhibiting Wnt/β-catenin signaling and reducing PD-L1 expression.DNASE1L3 overexpression similarly restricted organoid growth and suppressed pathway activity.Conclusion:DNASE1L3 acts as a negative regulator of HCC progression by targeting the Wnt/β-catenin pathway and reducing PD-L1 expression,thereby influencing both tumor cell behavior and macrophage-mediated immune responses.
基金supported by the Grant PID2021-126715OB-IOO financed by MCIN/AEI/10.13039/501100011033 and"ERDFA way of making Europe"by the Grant PI22CⅢ/00055 funded by Instituto de Salud CarlosⅢ(ISCⅢ)+6 种基金the UFIECPY 398/19(PEJ2018-004965) grant to RGS funded by AEI(Spain)the UFIECPY-396/19(PEJ2018-004961)grant financed by MCIN (Spain)FI23CⅢ/00003 grant funded by ISCⅢ-PFIS Spain) to PMMthe UFIECPY 328/22 (PEJ-2021-TL/BMD-21001) grant to LM financed by CAM (Spain)the grant by CAPES (Coordination for the Improvement of Higher Education Personnel)through the PDSE program (Programa de Doutorado Sanduiche no Exterior)to VSCG financed by MEC (Brazil)
文摘The brain is the most complex human organ,and commonly used models,such as two-dimensional-cell cultures and animal brains,often lack the sophistication needed to accurately use in research.In this context,human cerebral organoids have emerged as valuable tools offering a more complex,versatile,and human-relevant system than traditional animal models,which are often unable to replicate the intricate architecture and functionality of the human brain.Since human cerebral organoids are a state-of-the-art model for the study of neurodevelopment and different pathologies affecting the brain,this field is currently under constant development,and work in this area is abundant.In this review,we give a complete overview of human cerebral organoids technology,starting from the different types of protocols that exist to generate different human cerebral organoids.We continue with the use of brain organoids for the study of brain pathologies,highlighting neurodevelopmental,psychiatric,neurodegenerative,brain tumor,and infectious diseases.Because of the potential value of human cerebral organoids,we describe their use in transplantation,drug screening,and toxicology assays.We also discuss the technologies available to study cell diversity and physiological characteristics of organoids.Finally,we summarize the limitations that currently exist in the field,such as the development of vasculature and microglia,and highlight some of the novel approaches being pursued through bioengineering.
基金funded by the European Union’s Horizon 2020 research and innovation programme under the Marie Skodowska-Curie grant agreement No.813263(PMSMat Train,granted to UF,PP,MV,and DP)provided by the Fund for Scientific Research Flanders(FWO-Vlaanderen)of the Flemish Government(FWO sabbatical bench fee K800224N granted to PP)and ERA-NET Re Park(granted to PP)。
文摘Brain organoids encompass a large collection of in vitro stem cell–derived 3D culture systems that aim to recapitulate multiple aspects of in vivo brain development and function.First,this review provides a brief introduction to the current state-of-the-art for neuroectoderm brain organoid development,emphasizing their biggest advantages in comparison with classical two-dimensional cell cultures and animal models.However,despite their usefulness for developmental studies,a major limitation for most brain organoid models is the absence of contributing cell types from endodermal and mesodermal origin.As such,current research is highly investing towards the incorporation of a functional vasculature and the microglial immune component.In this review,we will specifically focus on the development of immune-competent brain organoids.By summarizing the different approaches applied to incorporate microglia,it is highlighted that immune-competent brain organoids are not only important for studying neuronal network formation,but also offer a clear future as a new tool to study inflammatory responses in vitro in 3D in a brainlike environment.Therefore,our main focus here is to provide a comprehensive overview of assays to measure microglial phenotype and function within brain organoids,with an outlook on how these findings could better understand neuronal network development or restoration,as well as the influence of physical stress on microglia-containing brain organoids.Finally,we would like to stress that even though the development of immune-competent brain organoids has largely evolved over the past decade,their full potential as a pre-clinical tool to study novel therapeutic approaches to halt or reduce inflammation-mediated neurodegeneration still needs to be explored and validated.
基金supported by the National Key Research and Development Program of China(No.2024YFA1300128)the National Natural Science Foundation of China(No.82372663)+2 种基金the Key Research and Development Program of Yunnan Province(No.202302AA310024)the Key Research and Development Program of Jiangxi Province(No.20232BBG70024)the Natural Science Foundation of Shandong Province(No.ZR2023LSW008).
文摘An organoid is a three-dimensional(3D)cell culture model that can reproduce the distinct structure and inherent functionality of certain organs.Nevertheless,a major limitation of organoids is the absence of a complex vascular network,thus restricting the supply of oxygen and essential nutrients.Coupled with their inherent size constraints and metabolite accumulation,it is challenging for organoids to replicate the natural intricacies of organs,thereby limiting their applicability.To overcome the challenges associated with this technology,we developed a culture platform to cultivate tumors or organ-derived organoids up to the centimeter scale.Initially,a customized organoid-on-a-chip including a microvascular network at the micron scale was designed using 3D printing.Further,by integrating an infusion device,the chip ensures an adequate supply of nutrients and fluid immersion while mimicking blood flow dynamics.Our method overcomes the issue of the limited size of organoids due to insufficient nutrient access,making it possible to produce large-scale tumor and normal tissue models in vitro,while providing insights into drug efficacy and toxicology evaluation as well as standardized organoid production.
基金funded by the National Natural Science Foundation of China(No.32300659)Shenzhen Science and Technology Innovation Commission Project(No.JCYJ20230807143302004)+6 种基金Zhangjiakou City Key R&D Plan Project(No.2421118D),Zhangjiakou City Key R&D Plan Project(No.2322088D)The Natural Science Project of Hebei North University(No.XJ2024034 and XJ2024035)Medical Science Research Subject Plan Project of Hebei Provincial Health Commission(No.20240782)Project of Administration of Traditional Chinese Medicine of Hebei Province(No.2025392)The 2025 Government-funded Training Project for Outstanding Clinical Medicine Talents(No.ZF2025264)Research Project of Medical Innovation for Chinese Youth(Project Leader:Jun Xue).g Project for Outstanding Clinical Medicine Talents(No.ZF2025264)Research Project of Medical Innovation for Chinese Youth(Project Leader:Jun Xue)。
文摘The structure of intestinal tissue is complex.In vitro simulation of intestinal structure and function is important for studying intestinal development and diseases.Recently,organoids have been successfully constructed and they have come to play an important role in biomedical research.Organoids are miniaturized three-dimensional(3D)organs,derived from stem cells,which mimic the structure,cell types,and physiological functions of an organ,making them robust models for biomedical research.Intestinal organoids are 3D micro-organs derived from intestinal stem cells or pluripotent stem cells that can successfully simulate the complex structure and function of the intestine,thereby providing a valuable platform for intestinal development and disease research.In this article,we review the latest progress in the construction and application of intestinal organoids.
基金supported by the National Natural Science Foundation of China(82372535 to Ru-Lin Huang and 82361138568 to Qingfeng Li)the Shanghai Clinical Research Center of Plastic and Reconstructive Surgery supported by Science and Technology Commission of Shanghai Municipality(22MC1940300)the Shanghai Plastic Surgery Research Center of Shanghai Priority Research Center(2023ZZ02023)。
文摘Current organoid-generation strategies rely predominantly on intricate in vitro manipulations of dissociated stem cells,including isolation,expansion,and genetic modification.However,these approaches present significant challenges in terms of safety and scalability for clinical applications.An alternative strategy involves the direct generation of organoids from readily available tissues.Herein,we report the generation of functional organoids representing all three germ layers from human adult adipose tissue without single-cell processing steps.Specifically,by employing a specialized suspension culture system,we have developed reaggregated microfat(RMF)tissues,which differentiated into mesodermal bone marrow organoids capable of reconstituting human normal hematopoiesis in immunodeficient mice,endodermal insulin-producing organoids that reversed hyperglycemia in streptozotocin(STZ)-induced diabetic mice,and ectodermal nervous-like tissues resembling neurons and neuroglial cells.These findings therefore highlight the potential of human adipose tissue as a safe,scalable,and clinically viable source for organoid-based regenerative therapies.
基金supported by the National Key Research and Development Program of China(2024YFA1108000 and 2021ZD0202500)the National Natural Science Foundation of China(32130035 and 92168107)+2 种基金the Joint Project of the Yangtze River Delta Science and Technology Innovation Community(2024CSJZN0600)the Central Guidance on Local Science and Technology Development Fund(YDZX20233100001002)the Shanghai Frontiers Science Center for Biomacromolecules and Precision Medicine at Shanghai Tech University.
文摘Hypoxic injury(HI)in the prenatal period often causes neonatal neurological disabilities.Due to the difficulty in obtaining clinical samples,the molecular and cellular mechanisms remain unclear.Here we use vascularized cerebral organoids to investigate the hypoxic injury phenotype and explore the intercellular interactions between vascular and neural tissues under hypoxic conditions.Our results indicate that fused vascularized cerebral organoids exhibit broader hypoxic responses and larger decreases in panels of neural development-related genes when exposed to low oxygen levels compared to single cerebral organoids.Interestingly,vessels also exhibit neural protective effects on T-box brain protein 2+intermediate progenitors(IPs),which are markedly lost in HI cerebral organoids.Furthermore,we identify the role of bone morphogenic protein signaling in protecting IPs.Thus,this study has established an in vitro organoid system that can be used to study the contribution of vessels to brain injury under hypoxic conditions and provides a strategy for the identification of intervention targets.
基金Westlake University and the Research Center for Industries of the Future of Westlake University,China(Grant No.:WU2022C040)authors acknowledge BioRender.com for the assistance of drawing Figs.1 and 4 and Graphical abstract.
文摘Brain organoid-on-chip platforms have emerged as groundbreaking tools in neural disease modeling and drug discovery,offering a unique and highly accurate simulation of human organ physiology and func-tion compared with traditional cell culture systems.This technology is a harmonious fusion of organ-on-a-chip and organoid culture technologies,leveraging their strengths to provide the most realistic in vitro replication of the in vivo environment,both physically and biologically.As both technologies continue to advance rapidly,this platform is highly promising in vitro platform for disease modeling.In this review,we summarize the historical developments,recent advancements,limitations,and future prospects of brain organoid-on-chip technology,aiming to illuminate the transformative potential of this platform in advancing our understanding and treatment of neural diseases.
基金supported by the National Key R&D Program of China,No.2019YFA0110300(to ZG)the National Natural Science Foundation of China,Nos.81773302(to YF),32070862(to ZG).
文摘Human brain development is a complex process,and animal models often have significant limitations.To address this,researchers have developed pluripotent stem cell-derived three-dimensional structures,known as brain-like organoids,to more accurately model early human brain development and disease.To enable more consistent and intuitive reproduction of early brain development,in this study,we incorporated forebrain organoid culture technology into the traditional unguided method of brain organoid culture.This involved embedding organoids in matrigel for only 7 days during the rapid expansion phase of the neural epithelium and then removing them from the matrigel for further cultivation,resulting in a new type of human brain organoid system.This cerebral organoid system replicated the temporospatial characteristics of early human brain development,including neuroepithelium derivation,neural progenitor cell production and maintenance,neuron differentiation and migration,and cortical layer patterning and formation,providing more consistent and reproducible organoids for developmental modeling and toxicology testing.As a proof of concept,we applied the heavy metal cadmium to this newly improved organoid system to test whether it could be used to evaluate the neurotoxicity of environmental toxins.Brain organoids exposed to cadmium for 7 or 14 days manifested severe damage and abnormalities in their neurodevelopmental patterns,including bursts of cortical cell death and premature differentiation.Cadmium exposure caused progressive depletion of neural progenitor cells and loss of organoid integrity,accompanied by compensatory cell proliferation at ectopic locations.The convenience,flexibility,and controllability of this newly developed organoid platform make it a powerful and affordable alternative to animal models for use in neurodevelopmental,neurological,and neurotoxicological studies.
基金supported by the Chinese Academy of Medical Sciences(Grant No.2021RU002)Beijing Natural Science Foundation(Grant No.Z240013)+2 种基金National Natural Science Foundation of China(Grant Nos.82450111,82388102,82373416,and 92259303)Beijing Research Ward Excellence Program(Grant Nos.BRWEP2024W034080200 and BRWEP2024W034080204)Peking University People’s Hospital Research and Development Funds(Grant No.RZG2024-02).
文摘Organoids are three-dimensional stem cell-derived models that offer a more physiologically relevant representation of tumor biology compared to traditional two-dimensional cell cultures or animal models.Organoids preserve the complex tissue architecture and cellular diversity of human cancers,enabling more accurate predictions of tumor growth,metastasis,and drug responses.Integration with microfluidic platforms,such as organ-on-a-chip systems,further enhances the ability to model tumor-environment interactions in real-time.Organoids facilitate in-depth exploration of tumor heterogeneity,molecular mechanisms,and the development of personalized treatment strategies when coupled with multi-omics technologies.Organoids provide a platform for investigating tumor-immune cell interactions,which aid in the design and testing of immune-based therapies and vaccines.Taken together,these features position organoids as a transformative tool in advancing cancer research and precision medicine.
基金supported by the Zhejiang Provincial Natural Science Foundation of China(LDT23H19012H19)the National Key R&D Program of China(2021YFC1712905)+5 种基金supported by the Innovation Team and Talents Cultivation Program of National Administration of Traditional Chinese Medicine(ZYYCXTD-D-202002)the Fundamental Research Funds for the Central Universities(226-2024-00001)Volker M.Lauschke was supported by the Swedish Research Council(2019-01837 and 2021-02801)Ruth och Richard Julins Foundation for Gastroenterology(2021-00158)the Knut and Alice Wallenberg Foundation(VC-2021-0026)Robert Bosch Foundation,Stuttgart,Germany.
文摘Coronavirus disease 2019(COVID-19),caused by severe acute respiratory syndrome coronavirus 2(SARS-CoV-2),not only affects the lungs but also damages various non-pulmonary organs,resulting in tissue injury and potential long-term sequelae in infected individuals.COVID-19 is likely to persist as a public health concern,given the frequent emergence of new mutations and viral strains.Multiple clinical lines of evidence indicate the efficacy of traditional Chinese medicine(TCM)in the prevention and treatment of COVID-19.However,the exact mechanism underlying these effects remains unclear.In this perspective review,we summarize the utility of in vitro three-dimensional(3D)cultured organoid models and organ-on-a-chip(OoC)technology for studying COVID-19 pathogenesis,viral tropism,and infectious mechanisms across different tissues.We highlight the successful application of these platforms in aiding drug development and discuss their advantages and limitations.We also review how such organotypic models can be employed to study TCMs.Finally,we discuss the opportunities for integrated microphysiological multi-tissue models to rapidly discover active components and potential targets in the context of COVID-19.The utilization of these emerging technologies could accelerate drug discovery and the modernization of TCM.
基金Supported by National Natural Science Foundation of China,No.81630080,No.82305179,and No.82374181The China Postdoctoral Science Foundation Grant,No.2019M650600+1 种基金The Beijing University of Chinese Medicine“Decoding Traditional Chinese Medicine”Project,No.2023-JYB-JBZD-036The Hefei National Research Center for Physical Sciences at the Microscale,No.KF2021104.
文摘BACKGROUND Hepatic organoid-based modelling,through the elucidation of a range of in vivo biological processes and the recreation of the intricate liver microenvironment,is yielding groundbreaking insights into the pathophysiology and personalized medicine approaches for liver diseases.AIM This study was designed to analyse the global scientific output of hepatic organoid research and assess current achievements and future trends through bibliometric analysis.METHODS Articles were retrieved from the Web of Science Core Collection,and CiteSpace 6.3.R1 was employed to analyse the literature,including outputs,journals,and countries,among others.RESULTS Between 2010 and 2024,a total of 991 articles pertaining to hepatic organoid research were published.The journal Hepatology published the greatest number of papers,and journals with an impact factor greater than 10 constituted 60%of the top 10 journals.The United States and Utrecht University were identified as the most prolific country and institution,respectively.Clevers H emerged as the most prolific author,whereas Huch M had the highest number of cocitations,suggesting that both are ideal candidates for academic collaboration.Research on hepatic organoids has exhibited a progressive shift in focus,evolving from initial investigations into model building,differentiation research in stem cells,bile ducts,and progenitor cells,to a broader spectrum encompassing lipid metabolism,single-cell RNA sequencing,and therapeutic applications.The phrases exhibiting citation bursts from 2022 to 2024 include“drug resistance”,“disease model”,and“patient-derived tumor organoids”.CONCLUSION Research on hepatic organoids has increased over the past decade and is expected to continue to grow.Key research areas include applications for liver diseases and drug development.Future trends likely to gain focus include patient-derived tumour organoids,disease modelling,and personalized medicine.
文摘Liposarcoma is one of the most common soft tissue sarcomas,however,its occurrence rate is still rare compared to other cancers.Due to its rarity,in vitro experiments are an essential approach to elucidate liposarcoma pathobiology.Conventional cell culture-based research(2D cell culture)is still playing a pivotal role,while several shortcomings have been recently under discussion.In vivo,mouse models are usually adopted for pre-clinical analyses with expectations to overcome the issues of 2D cell culture.However,they do not fully recapitulate human dedifferentiated liposarcoma(DDLPS)characteristics.Therefore,three-dimensional(3D)culture systems have been the recent research focus in the cell biology field with the expectation to overcome at the same time the disadvantages of 2D cell culture and in vivo animal models and fill in the gap between them.Given the liposarcoma rarity,we believe that 3D cell culture techniques,including 3D cell cultures/co-cultures,and Patient-Derived tumor Organoids(PDOs),represent a promising approach to facilitate liposarcoma investigation and elucidate its molecular mechanisms and effective therapy development.In this review,we first provide a general overview of 3D cell cultures compared to 2D cell cultures.We then focus on one of the recent 3D cell culture applications,Patient-Derived Organoids(PDOs),summarizing and discussing several PDO methodologies.Finally,we discuss the current and future applications of PDOs to sarcoma,particularly in the field of liposarcoma.
基金Supported by Conselho Nacional de Desenvolvimento Científico e Tecnológico,No.307318/2023-0 and No.102035/2024-5Fundação de AmparoàPesquisa do Estado de São Paulo,No.2023/10843-7 and No 2019/21070-3Nanotechnology National Laboratory System 2.0,Ministry of Science,Technology,Innovation and Communication,No.442539/2019-3.
文摘BACKGROUND Glioblastoma multiforme(GBM)is the most aggressive and prevalent primary malignant brain tumor in adults,marked by poor prognosis and high invasiveness.Traditional GBM invasion assays,such as those involving mouse brain xenografts,are often time-consuming and limited in efficiency.In this context,stem cell-derived neural organoids(NOs)have emerged as advanced,threedimensional,human-relevant platforms that mimic the cellular architecture and microenvironment of the human brain.These models provide novel opportunities to investigate glioblastoma stem cell invasion,a critical driver of tumor progression and therapeutic resistance.AIM To evaluate studies using stem cell-derived NOs to model glioblastoma migration/invasion,focusing on methodologies,applications and therapeutic implications.METHODS We conducted a systematic review following PRISMA guidelines,searching PubMed and Scopus for studies published between March 2019 and March 2025 that investigated NOs in the context of glioblastoma invasion/migration.After screening 377 articles based on predefined inclusion and exclusion criteria,10 original research articles were selected for analysis.Extracted data were categorized into four analytical domains:(1)Tumor model formation;(2)NO characteristics;(3)NO differentiation protocols;and(4)Invasion/migration assessment methodologies.RESULTS The included studies exhibit significant methodological heterogeneity GBM model development,particularly regarding model type,cell source and culture conditions.Most studies(70%)used suspension cell models,while 30%employed spheroids,with most research focusing on patient-derived glioblastoma stem cells.NOs were predominantly generated from human induced pluripotent stem cells using both guided and unguided differentiation protocols.Confocal fluorescence microscopy was the primary method used for assessing invasion,revealing invasion depths of up to 300μm.Organoid maturity and co-culture duration influenced results,while key factors for model optimization included tumor cell density,organoid age and extracellular matrix composition.Some studies also tested therapeutic strategies such as Zika virus and microRNA modulation.Collectively,findings support the utility of NOs as effective tools for studying GBM behavior and therapeutic responses in a humanized three-dimensional context.CONCLUSION Human NOs represent promising platforms for modeling glioblastoma invasion in a humanized three-dimensional environment.However,a limited number of studies and methodological heterogeneity hinder reproducibility.Protocol standardization is essential to enhance the translational application of these models.
基金supported by the National Natural Science Foundation of China(32201988)Natural Science Foundation of Jiangsu Province(BK20210456)+3 种基金Special Fund for Science and Technology Program of Jiangsu Province(BM2022019)the National Key R&D Program of China(2023YFC2506004)the Fundamental Research Funds for the Central Universities(JUSRP123047)the Program of Collaborative Innovation Centre of Food Safety and Quality Control in Jiangsu Province.
文摘The neurotransmitter 5-hydroxytryptamine(5-HT),primarily produced by intestinal enterochromaffin(EC)cells,relies on tryptophan hydroxylase 1(TPH1)for synthesis.Research suggested Bifidobacterium breve CCFM1025’s potential in regulating Tph1 gene expression,maintaining 5-HT levels in stressed mice,but its precise mechanisms were unclear.This study used metabolomic techniques to assess probiotic fermentation products,revealing acetate as the crucial element in Bb-CCFM1025’s regulation of gut 5-HT synthesis.Further exploration correlated acetate with Tph1 transcription in intestinal organoids.Transcriptomic methods and quantitative reverse transcription polymerase chain reaction validation demonstrated how acetate facilitated 5-HT synthesis and secretion.It unveiled that acetate orchestrates signaling pathways(phosphoinositide 3-kinase-protein kinase B(PI3K-AKT),phospholipase C-phosphorylated extracellular signal-regulated kinase(PLC-pERK),and PLC-1,4,5-trisphosphate(IP3)-Ca^(2+))within EC,enabling 5-HT production.These findings elucidate the biochemical mechanisms behind specific probiotics’effects,aiding in the targeted selection of similar beneficial strains.This study offers theoretical support for choosing probiotics with analogous functionalities based on their physiological impacts.
基金supported by the National Natural Science Foundation of China(Grant No.82074104)the National Natural Science Foundation of China(Grant No.82474169)the Research Project of Clinical Toxicology Transformation from the Chinese Society of Toxicology(No.CST2021CT101).
文摘Organoids are derived from self-organizing stem cells and form three-dimensional structures that are structurally and functionally similar to in vivo tissues.With the ability to replicate the in vivo microenvironment and maintain genetic stability,organoids have become a powerful tool for elucidating developmental mechanisms,accurately modeling disease processes,and efficiently screening drug candidates,and have also demonstrated significant value in the field of traditional Chinese medicine(TCM)-including applications in screening active components of TCM,studying TCM pharmacodynamic mechanisms,evaluating TCM safety,and verifying the effects of traditional non-pharmacological therapies such as acupuncture and yoga.Organoids can be cultured using air-liquid interface systems,bioreactors,and vascularization techniques.They are widely used in drug screening,disease modeling,precision medicine,and toxicity assessment.However,current limitations include high costs,difficulty in accurately replicating the microenvironment,and ethical concerns.In this review,we systematically retrieve,synthesize,and analyze relevant literature to elucidate the culture methods of organoid technology,its diverse applications across various fields,and the challenges it faces.In the future,integration with artificial intelligence may provide new insights and strategies for drug development and disease research and the modernization of TCM.
