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
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
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
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
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
Current experimental models struggle to simulate the complex process of the transformation from atrophic gastritis to gastric cancer,while gastric organoid technology,especially region-specific modeling,provides a mor...Current experimental models struggle to simulate the complex process of the transformation from atrophic gastritis to gastric cancer,while gastric organoid technology,especially region-specific modeling,provides a more precise in vitro platform for studying this carcinogenic mechanism.Helicobacter pylori activates carcinogenic signaling pathways through virulence factors,inducing DNA damage,epigenetic dysregulation,and immune microenvironment imbalance,driving inflammation-cancer conversion.Intestinal metaplasia and spasmolytic polypeptide-expressing metaplasia serve as critical precursor lesions,gradually developing into dysplasia and adenocarcinoma under the influence of chronic inflammation and genetic instability through intestinal cell transformation and high trefoil factor 2-expressing cell expansion.The immune suppression,metabolic reprogramming,and matrix remodeling within the tumor microenvironment collaboratively create a pro-cancer ecosystem that accelerates inflammationcarcinogenesis transformation.The gastric organoid model successfully simulates the spatiotemporal dynamics of the carcinogenesis process in atrophic gastritis,and its future integration with single-cell omics,real-time imaging technologies,and artificial intelligence technologies could provide a more precise platform for elucidating molecular mechanisms and screening intervention strategies.These advances position gastric organoids as pivotal tools for clinical translation,enabling personalized risk stratification,early intervention targeting precancerous transitions,and ex vivo prediction of patient-specific therapeutic responses to guide precision management of gastric cancer.展开更多
BACKGROUND Liver metastases are very common in pancreatic neuroendocrine tumors(pNETs).When surgical resection is possible,it is typically associated with survival benefits in patients with pNET and liver metastases.P...BACKGROUND Liver metastases are very common in pancreatic neuroendocrine tumors(pNETs).When surgical resection is possible,it is typically associated with survival benefits in patients with pNET and liver metastases.Patient-derived organoids are a powerful preclinical platform that show great potential for predicting treatment response,and they have been increasingly applied in precision medicine and cancer research.CASE SUMMARY A 51-year-old man was admitted to the hospital with the chief complaint of in-termittent dull pain in the upper abdomen for over 3 years.Computerized to-mography showed multiple space-occupying lesions in the liver and a neoplasm in the pancreatic body.Pathological results suggested a grade 3 pancreas-derived hepatic neuroendocrine tumor.In combination with relevant examinations,the patient was diagnosed with pNET with liver metastases(grade 3).Transarterial chemoembolization was initially performed with oxaliplatin and 5-fluorouracil,after which the chemotherapy regimen was switched to liposomal irinotecan and cisplatin for a subsequent perfusion,guided by organoid-based drug sensitivity testing.Following interventional treatment,the tumor had decreased in size.However,due to poor treatment compliance and the patient’s preference for sur-INTRODUCTION Pancreatic neuroendocrine tumors(pNETs)are a rare and heterogeneous group of neoplasms arising from pancreatic islet cells,with variations in histology,clinical characteristics,and prognosis[1].They may present as non-infiltrative,slow-growing tumors,locally invasive tumors,or even rapidly metastasizing tumors[2].Most metastases localize to the liver,and approximately 28%-77%of patients with pNETs will experience liver metastases in their lifetime[3].Patients with liver metastases may be subjected to local complications such as biliary obstruction,liver insufficiency,and carcinoid syndrome.Additionally,liver metastases are a major risk factor for the prognosis of patients with pNETs[4].When feasible,surgical resection is significantly associated with the best long-term survival outcomes[5].Therefore,for patients with pNET liver metastases,comprehensive assessment and multidisciplinary approaches are required to determine the feasibility of surgical resection and the optimal treatment to improve the prognosis.Over the past decade,the advent of in vitro three-dimensional technologies including organoids has revolutionized the development of human cancer models.Patient-derived organoids(PDOs),an in vitro three-dimensional microstructure,can faithfully recapitulate the intricate spatial architecture and cell heterogeneity of the tissue,and simulate the biological behaviors and functions of parental tumors while preserving biological,genetic and molecular features[6,7].As a po-werful preclinical platform,PDOs have been increasingly applied in precision medicine and cancer research.Importantly,there is a significant association between the use of PDO-based drug sensitivity testing and clinical responses to chemotherapy,radiotherapy and targeted therapy in multiple cancer types[8-10].Although gastroenteropancreatic neuroendocrine neoplasm organoids have been confirmed to retain the pathohistological and functional phenotypes of parental tumors[7],their roles in the prediction of clinical outcomes have not been presented.Here,we report a case of pNET with liver metastases who successfully received surgical resection after personalized treatment guided by PDO-based drug sensitivity testing,resulting in a favorable prognosis.展开更多
Major depressive disorder(MDD)is a debilitating psychiatric condition associated with substantial personal,societal,and economic costs.Despite considerable advances in research,most conventional antidepressant therapi...Major depressive disorder(MDD)is a debilitating psychiatric condition associated with substantial personal,societal,and economic costs.Despite considerable advances in research,most conventional antidepressant therapies fail to achieve adequate response in a significant proportion of patients,underscoring the need for novel,mechanism-based interventions.Lycium barbarum glycopeptide(LbGp),a bioactive compound with emerging neuroprotective properties,has been proposed as a candidate for antidepressant development;however,its therapeutic efficacy and underlying mechanisms remain largely uncharacterized.In this study,ventral forebrain organoids were generated from patients with MDD to investigate disease-related neurophysiological abnormalities.These organoids exhibited disrupted neuronal morphology,diminished calcium signaling,and impaired electrophysiological activity.Administration of LbGp effectively restored structural and functional deficits in MDD-derived organoids.Transcriptomic profiling revealed that LbGp ameliorated endoplasmic reticulum(ER)stress responses.To investigate the causative role of ER stress,control organoids were treated with the ER stress agonist CCT020312,which elicited neural activity impairments resembling those observed in MDD organoids.Notably,LbGp reversed the phenotypic consequences of CCT020312 exposure in control organoids.In conclusion,ventral forebrain organoids derived from individuals with MDD demonstrated that LbGp ameliorates disease-associated phenotypes by modulating ER stress.展开更多
AIM:To explore the changes in early retinal development after the occurrence of ischemia.METHODS:Human retinal organoids(hROs)of day 18 or day 30 were treated with oxygen-glucose deprivation and reperfusion(OGD/R)to s...AIM:To explore the changes in early retinal development after the occurrence of ischemia.METHODS:Human retinal organoids(hROs)of day 18 or day 30 were treated with oxygen-glucose deprivation and reperfusion(OGD/R)to simulate the retinal ischemia.All hROs were maintained normally until day 60 to evaluate changes in ischemic injuries during retinal development.Paraffin section staining was used for detecting changes in organoid structure and cell number.Real-time quantitative polymerase chain reaction(RT-qPCR)and Western blot(WB)analyses were used to observe the change in the expression of retinal cell markers.RESULTS:In hROs,OGD/R induced the decrease of proliferating cells,inhibited the expression of proliferated marker Ki67 and promoted early apoptosis of retinal cells(P<0.05).Under OGD/R condition,the progenitor cell layer and ganglion cell layer of hROs lost normal structure,and the number of neural stem cells(SOX2^(+)),retinal progenitor cells(CHX10^(+))and retinal ganglion cells(TUJ1^(+)/BRN3^(+)/ATOH7^(+))decreased(P<0.05).The expression of corresponding retinal cell markers also decreased(P<0.05).Organoids treated with OGD/R on day 30 had similar injuries in retinal structure and retinal cell markers to those on day 18.Long-term observations revealed that day 18-treated organoids remained disorganized progenitor and ganglion cell layers by day 60,with no recovery in proliferating cell nuclear antigen(PCNA)protein expression.RT-qPCR showed persistently low Ki67 transcription levels(P<0.001),while other retinal cell markers recovered or exceeded normal levels,indicating a limited self-repair happened in the development of hROs.In contrast,day 30-treated organoids exhibited normal structure and marker expression by day 60,with transcription levels of retinal cell markers returning to normal(P>0.05),demonstrating complete recovery from OGD/R damage.CONCLUSION:Retinal ischemia damage the retinal development in the short-term.After the restoration of retinal blood supply,the retinal ischemic damage can be recovered during subsequent development.However,retinal ischemic injuries at different developmental stages exhibit varying degrees of reversibility.The earlier ischemic injury occurs,the more difficult it is to repair retinal cell and structure damage.展开更多
Hepatobiliary and pancreatic(HBP)cancers are among the most aggressive malig-nancies,with recurrence and metastasis driven by tumor heterogeneity and drug resistance,presenting considerable challenges to effective tre...Hepatobiliary and pancreatic(HBP)cancers are among the most aggressive malig-nancies,with recurrence and metastasis driven by tumor heterogeneity and drug resistance,presenting considerable challenges to effective treatment.Currently,personalized and accurate treatment prediction models for these cancers are lacking.Patient-derived organoids(PDOs)tumor are three-dimensional in vitro models created from the tumor tissues of individual patients.Recent reports and our cultivation data indicate that the success rate of cultivating organoids for HBP cancers consistently exceeds 70%.The predictive accuracy of these tumor orga-noids has been shown to surpass 90%.However,PDOs still face notable limita-tions,especially in simulating the tumor microenvironment,including tumor angiogenesis and the surrounding cellular context,which require further refi-nement.While co-culture techniques and microfluidic platforms have been deve-loped to mimic multi-cellular environments and functional vascular perfusion,they remain insufficient in accurately recapitulating the complexities of the in vivo environment.Additionally,PDOs are needed to fully assess their potential in predicting the efficacy of multi-drug combination therapies.This review provides an overview of the applications,challenges,and prospects for organoid models in the study of HBP cancer.展开更多
Organoids are three-dimensional structures derived from stem cells that recapitulate the gene expression profiles and functional characteristics of their tissue of origin,rendering them invaluable tools for disease mo...Organoids are three-dimensional structures derived from stem cells that recapitulate the gene expression profiles and functional characteristics of their tissue of origin,rendering them invaluable tools for disease modeling,drug screening,and precision medicine.Despite their promise,the widespread application of organoids is limited by extended culture durations and technical complexity.Cryopreservation has emerged as a critical strategy to overcome challenges related to the long-term storage and application of organoids,offering a range of preservation approaches tailored to organoid development.Nevertheless,conventional cryopreservation techniques encounter significant limitations when applied to organoids.To address these issues,the development of naturally derived,low-toxicity Cryoprotectants(CPAs),along with the optimization of CPA loading methods and refinement of cooling and warming protocols,is essential to mitigate cryoinjury.Looking forward,the comprehensive enhancement of cryopreservation technologies may facilitate the transformation of organoids into“off-the-shelf”products,enabling scalable production,batch standardization,and centralized distribution.Such advancements will lay the foundation for the establishment of Next-Generation Living Biobanks(NGLB).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
BACKGROUND Human-derived gastric cancer organoids(GCOs)are widely used in gastric cancer research;however,the culture success rate is generally low.AIM To explore the potential influencing factors,and the literature o...BACKGROUND Human-derived gastric cancer organoids(GCOs)are widely used in gastric cancer research;however,the culture success rate is generally low.AIM To explore the potential influencing factors,and the literature on successful culture rates of GCOs was reviewed using meta-analysis.METHODS PubMed,Web of Science,and EMBASE were searched for studies.Two trained researchers selected the studies and extracted data.STATA 17.0 software was used for meta-analysis of the incidence of each outcome event.The adjusted Methodological Index for Non-Randomized Studies scale was used to assess the quality of the included studies.Funnel plots and Egger’s test were used to detect publication bias.Subgroup analyses were conducted for sex,tissue source,histo-logical classification,and the pathological tumor-node-metastasis(pTNM)cancer staging system.RESULTS Eight studies with a pooled success rate of 66.6%were included.GCOs derived from women and men had success rates of 67%and 46.7%,respectively.GCOs from surgery or biopsy/endoscopic submucosal dissection showed success rates of 70.9%and 53.7%,respectively.GCOs of poorly-differentiated,moderately-differentiated and signet-ring cell cancer showed success rates of 64.6%,31%,and 32.7%,respectively.GCOs with pTNM stages I-II and III-IV showed success rates of 38.3%and 65.2%,respectively.Y-27632 and non-Y-27632 use showed success rates of 58.2%and 70%,respectively.GCOs generated with collagenase were more successful than those constructed with Liberase TH and TrypLE(72.1%vs 71%,respectively).EDTA digestion showed a 50%lower success rate than other methods(P=0.04).CONCLUSION GCO establishment rate is low and varies by sex,tissue source,histological type,and pTNM stage.Omitting Y-27632,and using Liberase TH,TrypLE,or collagenase yields greater success than EDTA.展开更多
The liver,the largest solid organ in the body,is susceptible to metabolic diseases and malignant tumors.Studying its physiological and pathological processes helps to optimize the clinical treatment.Organoids are a no...The liver,the largest solid organ in the body,is susceptible to metabolic diseases and malignant tumors.Studying its physiological and pathological processes helps to optimize the clinical treatment.Organoids are a novel tool for studying physical development,disease mechanisms,and high-throughput drug screening due to their similarity in composition,structure,and function to internal organs.Recent studies have shown that stem cells,hepatocytes,or cholangiocytes can form“liver organoids”under the synergistic action of specific extracellular matrix and various signaling molecules.This review outlines techniques for generating liver organoids that maximally recapitulate the liver structure and functions in vitro and thoroughly discusses the customary applications of organoids derived from liver tissue,induced pluripotent stem cells(iPSCs)and liver tumors.In this review,a meticulous analysis is provided of the comparatively advanced culture systems used in the construction of liver cancer-derived organoids.Additionally,we reviewed the progress of liver organoids in disease modeling,drug efficacy,and toxicity evaluation,in hopes of generating innovative ideas for the research and applications of liver organoids.展开更多
Spinal cord organoids are three-dimensional tissues derived from stem cells that recapitulate the primary morphological and functional characteristics of the spinal cord in vivo.As emerging bioengineering methods have...Spinal cord organoids are three-dimensional tissues derived from stem cells that recapitulate the primary morphological and functional characteristics of the spinal cord in vivo.As emerging bioengineering methods have led to the optimization of cell culture protocols,spinal cord organoids technology has made remarkable advancements in the past decade.Our literature search found that current spinal cord organoids do not only dynamically simulate neural tube formation but also exhibit diverse cytoarchitecture along the dorsal-ventral and rostral-caudal axes.Moreover,fused organoids that integrate motor neurons and other regionally specific organoids exhibit intricate neural circuits that allows for functional assessment.These qualities make spinal cord organoids valuable tools for disease modeling,drug screening,and tissue regeneration.By utilizing this emergent technology,researchers have made significant progress in investigating the pathogenesis and potential therapeutic targets of spinal cord diseases.However,at present,spinal cord organoid technology remains in its infancy and has not been widely applied in translational medicine.Establishment of the next generation of spinal cord organoids will depend on good manufacturing practice standards and needs to focus on diverse cell phenotypes and electrophysiological functionality evaluation.展开更多
Human pluripotent stem cell(hPSC)-derived kidney organoids share similarities with the fetal kidney.However,the current hPSC-derived kidney organoids have some limitations,including the inability to perform nephrogene...Human pluripotent stem cell(hPSC)-derived kidney organoids share similarities with the fetal kidney.However,the current hPSC-derived kidney organoids have some limitations,including the inability to perform nephrogenesis and lack of a corticomedullary definition,uniform vascular system,and coordinated exit path-way for urinary filtrate.Therefore,further studies are required to produce hPSC-derived kidney organoids that accurately mimic human kidneys to facilitate research on kidney development,regeneration,disease modeling,and drug screening.In this review,we discussed recent advances in the generation of hPSC-derived kidney organoids,how these organoids contribute to the understanding of human kidney development and research in disease modeling.Additionally,the limitations,future research focus,and applications of hPSC-derived kidney organoids were highlighted.展开更多
Neural tube defects(NTDs)are severe congenital neurodevelopmental disorders arising from incomplete neural tube closure.Although folate supplementation has been shown to mitigate the incidence of NTDs,some cases,often...Neural tube defects(NTDs)are severe congenital neurodevelopmental disorders arising from incomplete neural tube closure.Although folate supplementation has been shown to mitigate the incidence of NTDs,some cases,often attributable to genetic factors,remain unpreventable.The SHROOM3 gene has been implicated in NTD cases that are unresponsive to folate supplementation;at present,however,the underlying mechanism remains unclear.Neural tube morphogenesis is a complex process involving the folding of the planar epithelium of the neural plate.To determine the role of SHROOM3 in early developmental morphogenesis,we established a neuroepithelial organoid culture system derived from cynomolgus monkeys to closely mimic the in vivo neural plate phase.Loss of SHROOM3 resulted in shorter neuroepithelial cells and smaller nuclei.These morphological changes were attributed to the insufficient recruitment of cytoskeletal proteins,namely fibrous actin(F-actin),myosin II,and phospho-myosin light chain(PMLC),to the apical side of the neuroepithelial cells.Notably,these defects were not rescued by folate supplementation.RNA sequencing revealed that differentially expressed genes were enriched in biological processes associated with cellular and organ morphogenesis.In summary,we established an authentic in vitro system to study NTDs and identified a novel mechanism for NTDs that are unresponsive to folate supplementation.展开更多
Lung cancer is emerging as a common malignancy worldwide,with non-small cell lung cancer(NSCLC)accounting for approximately 85%of all cases.Two-dimensional(2D)in vitro cell line cultures and animal models are currentl...Lung cancer is emerging as a common malignancy worldwide,with non-small cell lung cancer(NSCLC)accounting for approximately 85%of all cases.Two-dimensional(2D)in vitro cell line cultures and animal models are currently used to study NSCLC.However,2D cell cultures fail to replicate the medication response and neoplastic heterogeneity of parental tumors.Animal models are expensive and require lengthy modeling cycles.The generation of in vitro three-dimensional(3D)tissue cultures called organoids,which exhibit multicellular,anatomical,and functional properties of real organs,is now achievable owing to advancements in stem cell culturing.The genetic,proteomic,morphological,and pharmacological characteristics of tumors are largely preserved in tumor organoids grown in vitro.The design and physiology of human organs can be precisely reconstructed in tumor organoids,opening new possibilities for complementing the use of animal models and studying human diseases.This review summarizes the development of NSCLC organoids and their applications in basic research,drug testing,immunotherapy,and individualized treatments.展开更多
Gastric organoids are models created in the laboratory using stem cells and sophisticated three-dimensional cell culture techniques.These models have shown great promise in providing valuable insights into gastric phy...Gastric organoids are models created in the laboratory using stem cells and sophisticated three-dimensional cell culture techniques.These models have shown great promise in providing valuable insights into gastric physiology and advanced disease research.This review comprehensively summarizes and analyzes the research advances in culture methods and techniques for adult stem cells and induced pluripotent stem cell-derived organoids,and patient-derived organoids.The potential value of gastric organoids in studying the pathogenesis of stomach-related diseases and facilitating drug screening is initially discussed.The construction of gastric organoids involves several key steps,including cell extraction and culture,three-dimensional structure formation,and functional expression.Simulating the structure and function of the human stomach by disease modeling with gastric organoids provides a platform to study the mechanism of gastric cancer induction by Helicobacter pylori.In addition,in drug screening and development,gastric organoids can be used as a key tool to evaluate drug efficacy and toxicity in preclinical trials.They can also be used for precision medicine according to the specific conditions of patients with gastric cancer,to assess drug resistance,and to predict the possibility of adverse reactions.However,despite the impressive progress in the field of gastric organoids,there are still many unknowns that need to be addressed,especially in the field of regenerative medicine.Meanwhile,the reproducibility and consistency of organoid cultures are major challenges that must be overcome.These challenges have had a significant impact on the development of gastric organoids.Nonetheless,as technology continues to advance,we can foresee more comprehensive research in the construction of gastric organoids.Such research will provide better solutions for the treatment of stomach-related diseases and personalized medicine.展开更多
BACKGROUND Patient-derived organoids(PDOs)have been demonstrated to predict the response to drugs in multiple cancer types.However,it remains unclear about its application in cholangiocarcinoma.CASE SUMMARY A 59-year-...BACKGROUND Patient-derived organoids(PDOs)have been demonstrated to predict the response to drugs in multiple cancer types.However,it remains unclear about its application in cholangiocarcinoma.CASE SUMMARY A 59-year-old woman was admitted to the hospital due to upper abdominal pain for over 8 months.According to relevant examinations,she was diagnosed as perihilar cholangiocarcinoma(pCCA)with intrahepatic metastasis and perihilar lymphatic metastasis.After multidisciplinary team discussion,percutaneous transhepatic cholangiodrainage was performed to relieve biliary obstruction,and puncture biopsy was conducted to confirm the pathological diagnosis.Transarterial chemoembolization with nab-paclitaxel was used in combination with toripalimab and lenvatinib,but the levels of tumor markers including alpha fetal protein,carcinoembryonic antigen,carbohydrate antigen 15-3 and cancer antigen 125 were still raised.The PDO for drug screening showed sensitive to gemcitabine and cisplatin.Accordingly,the chemotherapy regimen was adjusted to gemcitabine and cisplatin in combination with toripalimab and lenvatinib.After 4 cycles of treatment,the tumor was assessed resectable,and radical surgical resection was performed successfully.One year after surgery,the patient was still alive,and no recurrence or occurred.CONCLUSION PDOs for drug sensitivity contribute to screening effective chemotherapy drugs for advanced pCCA,promoting conversion therapy and improving the prognosis.展开更多
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 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 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 National Traditional Chinese Medicine Advantageous Specialty Project of National Administration of Traditional Chinese MedicineShuguang Hospital Siming Foundation Research Special Project,No.SGKJ-202304.
文摘Current experimental models struggle to simulate the complex process of the transformation from atrophic gastritis to gastric cancer,while gastric organoid technology,especially region-specific modeling,provides a more precise in vitro platform for studying this carcinogenic mechanism.Helicobacter pylori activates carcinogenic signaling pathways through virulence factors,inducing DNA damage,epigenetic dysregulation,and immune microenvironment imbalance,driving inflammation-cancer conversion.Intestinal metaplasia and spasmolytic polypeptide-expressing metaplasia serve as critical precursor lesions,gradually developing into dysplasia and adenocarcinoma under the influence of chronic inflammation and genetic instability through intestinal cell transformation and high trefoil factor 2-expressing cell expansion.The immune suppression,metabolic reprogramming,and matrix remodeling within the tumor microenvironment collaboratively create a pro-cancer ecosystem that accelerates inflammationcarcinogenesis transformation.The gastric organoid model successfully simulates the spatiotemporal dynamics of the carcinogenesis process in atrophic gastritis,and its future integration with single-cell omics,real-time imaging technologies,and artificial intelligence technologies could provide a more precise platform for elucidating molecular mechanisms and screening intervention strategies.These advances position gastric organoids as pivotal tools for clinical translation,enabling personalized risk stratification,early intervention targeting precancerous transitions,and ex vivo prediction of patient-specific therapeutic responses to guide precision management of gastric cancer.
基金Supported by Chongqing Natural Science Foundation General Project,No.CSTB2023NSCQ-MSX0182 and No.CSTB2023NSCQMSX0252Clinical Research Special Project of The Second Affiliated Hospital of Army Medical University,No.2024 F022.
文摘BACKGROUND Liver metastases are very common in pancreatic neuroendocrine tumors(pNETs).When surgical resection is possible,it is typically associated with survival benefits in patients with pNET and liver metastases.Patient-derived organoids are a powerful preclinical platform that show great potential for predicting treatment response,and they have been increasingly applied in precision medicine and cancer research.CASE SUMMARY A 51-year-old man was admitted to the hospital with the chief complaint of in-termittent dull pain in the upper abdomen for over 3 years.Computerized to-mography showed multiple space-occupying lesions in the liver and a neoplasm in the pancreatic body.Pathological results suggested a grade 3 pancreas-derived hepatic neuroendocrine tumor.In combination with relevant examinations,the patient was diagnosed with pNET with liver metastases(grade 3).Transarterial chemoembolization was initially performed with oxaliplatin and 5-fluorouracil,after which the chemotherapy regimen was switched to liposomal irinotecan and cisplatin for a subsequent perfusion,guided by organoid-based drug sensitivity testing.Following interventional treatment,the tumor had decreased in size.However,due to poor treatment compliance and the patient’s preference for sur-INTRODUCTION Pancreatic neuroendocrine tumors(pNETs)are a rare and heterogeneous group of neoplasms arising from pancreatic islet cells,with variations in histology,clinical characteristics,and prognosis[1].They may present as non-infiltrative,slow-growing tumors,locally invasive tumors,or even rapidly metastasizing tumors[2].Most metastases localize to the liver,and approximately 28%-77%of patients with pNETs will experience liver metastases in their lifetime[3].Patients with liver metastases may be subjected to local complications such as biliary obstruction,liver insufficiency,and carcinoid syndrome.Additionally,liver metastases are a major risk factor for the prognosis of patients with pNETs[4].When feasible,surgical resection is significantly associated with the best long-term survival outcomes[5].Therefore,for patients with pNET liver metastases,comprehensive assessment and multidisciplinary approaches are required to determine the feasibility of surgical resection and the optimal treatment to improve the prognosis.Over the past decade,the advent of in vitro three-dimensional technologies including organoids has revolutionized the development of human cancer models.Patient-derived organoids(PDOs),an in vitro three-dimensional microstructure,can faithfully recapitulate the intricate spatial architecture and cell heterogeneity of the tissue,and simulate the biological behaviors and functions of parental tumors while preserving biological,genetic and molecular features[6,7].As a po-werful preclinical platform,PDOs have been increasingly applied in precision medicine and cancer research.Importantly,there is a significant association between the use of PDO-based drug sensitivity testing and clinical responses to chemotherapy,radiotherapy and targeted therapy in multiple cancer types[8-10].Although gastroenteropancreatic neuroendocrine neoplasm organoids have been confirmed to retain the pathohistological and functional phenotypes of parental tumors[7],their roles in the prediction of clinical outcomes have not been presented.Here,we report a case of pNET with liver metastases who successfully received surgical resection after personalized treatment guided by PDO-based drug sensitivity testing,resulting in a favorable prognosis.
基金supported by the National Key Research and Development Program of China(2021YFA1101800,2022YFA1104800,2023YFF1203600)National Natural Science Foundation of China(82325015,82171528,U23A20429,82371260,21904069,22274079,82401794)+2 种基金Jiangsu Provincial Natural Science Fund for Excellent Young Scholars(BK20240131)Natural Science Foundation of Jiangsu Province(BK20200677)Joint Project of the Yangtze River Delta Science and Technology Innovation Community(2024CSJZN0600)。
文摘Major depressive disorder(MDD)is a debilitating psychiatric condition associated with substantial personal,societal,and economic costs.Despite considerable advances in research,most conventional antidepressant therapies fail to achieve adequate response in a significant proportion of patients,underscoring the need for novel,mechanism-based interventions.Lycium barbarum glycopeptide(LbGp),a bioactive compound with emerging neuroprotective properties,has been proposed as a candidate for antidepressant development;however,its therapeutic efficacy and underlying mechanisms remain largely uncharacterized.In this study,ventral forebrain organoids were generated from patients with MDD to investigate disease-related neurophysiological abnormalities.These organoids exhibited disrupted neuronal morphology,diminished calcium signaling,and impaired electrophysiological activity.Administration of LbGp effectively restored structural and functional deficits in MDD-derived organoids.Transcriptomic profiling revealed that LbGp ameliorated endoplasmic reticulum(ER)stress responses.To investigate the causative role of ER stress,control organoids were treated with the ER stress agonist CCT020312,which elicited neural activity impairments resembling those observed in MDD organoids.Notably,LbGp reversed the phenotypic consequences of CCT020312 exposure in control organoids.In conclusion,ventral forebrain organoids derived from individuals with MDD demonstrated that LbGp ameliorates disease-associated phenotypes by modulating ER stress.
基金Supported by the National Natural Science Foundation of China(No.82070937).
文摘AIM:To explore the changes in early retinal development after the occurrence of ischemia.METHODS:Human retinal organoids(hROs)of day 18 or day 30 were treated with oxygen-glucose deprivation and reperfusion(OGD/R)to simulate the retinal ischemia.All hROs were maintained normally until day 60 to evaluate changes in ischemic injuries during retinal development.Paraffin section staining was used for detecting changes in organoid structure and cell number.Real-time quantitative polymerase chain reaction(RT-qPCR)and Western blot(WB)analyses were used to observe the change in the expression of retinal cell markers.RESULTS:In hROs,OGD/R induced the decrease of proliferating cells,inhibited the expression of proliferated marker Ki67 and promoted early apoptosis of retinal cells(P<0.05).Under OGD/R condition,the progenitor cell layer and ganglion cell layer of hROs lost normal structure,and the number of neural stem cells(SOX2^(+)),retinal progenitor cells(CHX10^(+))and retinal ganglion cells(TUJ1^(+)/BRN3^(+)/ATOH7^(+))decreased(P<0.05).The expression of corresponding retinal cell markers also decreased(P<0.05).Organoids treated with OGD/R on day 30 had similar injuries in retinal structure and retinal cell markers to those on day 18.Long-term observations revealed that day 18-treated organoids remained disorganized progenitor and ganglion cell layers by day 60,with no recovery in proliferating cell nuclear antigen(PCNA)protein expression.RT-qPCR showed persistently low Ki67 transcription levels(P<0.001),while other retinal cell markers recovered or exceeded normal levels,indicating a limited self-repair happened in the development of hROs.In contrast,day 30-treated organoids exhibited normal structure and marker expression by day 60,with transcription levels of retinal cell markers returning to normal(P>0.05),demonstrating complete recovery from OGD/R damage.CONCLUSION:Retinal ischemia damage the retinal development in the short-term.After the restoration of retinal blood supply,the retinal ischemic damage can be recovered during subsequent development.However,retinal ischemic injuries at different developmental stages exhibit varying degrees of reversibility.The earlier ischemic injury occurs,the more difficult it is to repair retinal cell and structure damage.
基金Supported by Key Research and Development Program of Zhejiang Province,No.2024C03172Clinical Medical Research Special Fund Project of Zhejiang Medical Association,No.2024ZYC-Z50.
文摘Hepatobiliary and pancreatic(HBP)cancers are among the most aggressive malig-nancies,with recurrence and metastasis driven by tumor heterogeneity and drug resistance,presenting considerable challenges to effective treatment.Currently,personalized and accurate treatment prediction models for these cancers are lacking.Patient-derived organoids(PDOs)tumor are three-dimensional in vitro models created from the tumor tissues of individual patients.Recent reports and our cultivation data indicate that the success rate of cultivating organoids for HBP cancers consistently exceeds 70%.The predictive accuracy of these tumor orga-noids has been shown to surpass 90%.However,PDOs still face notable limita-tions,especially in simulating the tumor microenvironment,including tumor angiogenesis and the surrounding cellular context,which require further refi-nement.While co-culture techniques and microfluidic platforms have been deve-loped to mimic multi-cellular environments and functional vascular perfusion,they remain insufficient in accurately recapitulating the complexities of the in vivo environment.Additionally,PDOs are needed to fully assess their potential in predicting the efficacy of multi-drug combination therapies.This review provides an overview of the applications,challenges,and prospects for organoid models in the study of HBP cancer.
基金supported by the National Natural Science Foundation of China(52076140).
文摘Organoids are three-dimensional structures derived from stem cells that recapitulate the gene expression profiles and functional characteristics of their tissue of origin,rendering them invaluable tools for disease modeling,drug screening,and precision medicine.Despite their promise,the widespread application of organoids is limited by extended culture durations and technical complexity.Cryopreservation has emerged as a critical strategy to overcome challenges related to the long-term storage and application of organoids,offering a range of preservation approaches tailored to organoid development.Nevertheless,conventional cryopreservation techniques encounter significant limitations when applied to organoids.To address these issues,the development of naturally derived,low-toxicity Cryoprotectants(CPAs),along with the optimization of CPA loading methods and refinement of cooling and warming protocols,is essential to mitigate cryoinjury.Looking forward,the comprehensive enhancement of cryopreservation technologies may facilitate the transformation of organoids into“off-the-shelf”products,enabling scalable production,batch standardization,and centralized distribution.Such advancements will lay the foundation for the establishment of Next-Generation Living Biobanks(NGLB).
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 National Natural Science Foundation of China,No.82174309 and No.81973774National Administration of Traditional Chinese Medicine:2019 Project of Building Evidence-Based Practice Capacity for TCM,No.2019XZZX-XH013Shuguang Hospital Siming Foundation Research Special Project,No.SGKJ-202304.
文摘BACKGROUND Human-derived gastric cancer organoids(GCOs)are widely used in gastric cancer research;however,the culture success rate is generally low.AIM To explore the potential influencing factors,and the literature on successful culture rates of GCOs was reviewed using meta-analysis.METHODS PubMed,Web of Science,and EMBASE were searched for studies.Two trained researchers selected the studies and extracted data.STATA 17.0 software was used for meta-analysis of the incidence of each outcome event.The adjusted Methodological Index for Non-Randomized Studies scale was used to assess the quality of the included studies.Funnel plots and Egger’s test were used to detect publication bias.Subgroup analyses were conducted for sex,tissue source,histo-logical classification,and the pathological tumor-node-metastasis(pTNM)cancer staging system.RESULTS Eight studies with a pooled success rate of 66.6%were included.GCOs derived from women and men had success rates of 67%and 46.7%,respectively.GCOs from surgery or biopsy/endoscopic submucosal dissection showed success rates of 70.9%and 53.7%,respectively.GCOs of poorly-differentiated,moderately-differentiated and signet-ring cell cancer showed success rates of 64.6%,31%,and 32.7%,respectively.GCOs with pTNM stages I-II and III-IV showed success rates of 38.3%and 65.2%,respectively.Y-27632 and non-Y-27632 use showed success rates of 58.2%and 70%,respectively.GCOs generated with collagenase were more successful than those constructed with Liberase TH and TrypLE(72.1%vs 71%,respectively).EDTA digestion showed a 50%lower success rate than other methods(P=0.04).CONCLUSION GCO establishment rate is low and varies by sex,tissue source,histological type,and pTNM stage.Omitting Y-27632,and using Liberase TH,TrypLE,or collagenase yields greater success than EDTA.
基金supported by Research Projects from the Science and Technology Commission of Shanghai Municipality(No.21JC1401200).
文摘The liver,the largest solid organ in the body,is susceptible to metabolic diseases and malignant tumors.Studying its physiological and pathological processes helps to optimize the clinical treatment.Organoids are a novel tool for studying physical development,disease mechanisms,and high-throughput drug screening due to their similarity in composition,structure,and function to internal organs.Recent studies have shown that stem cells,hepatocytes,or cholangiocytes can form“liver organoids”under the synergistic action of specific extracellular matrix and various signaling molecules.This review outlines techniques for generating liver organoids that maximally recapitulate the liver structure and functions in vitro and thoroughly discusses the customary applications of organoids derived from liver tissue,induced pluripotent stem cells(iPSCs)and liver tumors.In this review,a meticulous analysis is provided of the comparatively advanced culture systems used in the construction of liver cancer-derived organoids.Additionally,we reviewed the progress of liver organoids in disease modeling,drug efficacy,and toxicity evaluation,in hopes of generating innovative ideas for the research and applications of liver organoids.
基金supported by the sup-project of National Key R&D Program of China,No.2018YFA0108602CAMS Innovation Fund for Medical Sciences,No.CIFMS,2021-I2M-C&T-B-016National High Level Hospital Clinical Research Funding,No.2022-PUMCH-B-112(all to JG).
文摘Spinal cord organoids are three-dimensional tissues derived from stem cells that recapitulate the primary morphological and functional characteristics of the spinal cord in vivo.As emerging bioengineering methods have led to the optimization of cell culture protocols,spinal cord organoids technology has made remarkable advancements in the past decade.Our literature search found that current spinal cord organoids do not only dynamically simulate neural tube formation but also exhibit diverse cytoarchitecture along the dorsal-ventral and rostral-caudal axes.Moreover,fused organoids that integrate motor neurons and other regionally specific organoids exhibit intricate neural circuits that allows for functional assessment.These qualities make spinal cord organoids valuable tools for disease modeling,drug screening,and tissue regeneration.By utilizing this emergent technology,researchers have made significant progress in investigating the pathogenesis and potential therapeutic targets of spinal cord diseases.However,at present,spinal cord organoid technology remains in its infancy and has not been widely applied in translational medicine.Establishment of the next generation of spinal cord organoids will depend on good manufacturing practice standards and needs to focus on diverse cell phenotypes and electrophysiological functionality evaluation.
基金the National Natural Science Foundation of China,No.82360148Guizhou Science&Technology Department,No.QKHPTRC2018-5636-2 and No.QKHPTRC2020-2201.
文摘Human pluripotent stem cell(hPSC)-derived kidney organoids share similarities with the fetal kidney.However,the current hPSC-derived kidney organoids have some limitations,including the inability to perform nephrogenesis and lack of a corticomedullary definition,uniform vascular system,and coordinated exit path-way for urinary filtrate.Therefore,further studies are required to produce hPSC-derived kidney organoids that accurately mimic human kidneys to facilitate research on kidney development,regeneration,disease modeling,and drug screening.In this review,we discussed recent advances in the generation of hPSC-derived kidney organoids,how these organoids contribute to the understanding of human kidney development and research in disease modeling.Additionally,the limitations,future research focus,and applications of hPSC-derived kidney organoids were highlighted.
基金supported by the National Natural Science Foundation of China (81930121,82125008 to Y.C.C.)National Key Research and Development Program of China (2018YFA0107902 to Y.C.C.and 2018YFA0801403 to Z.B.W.)+1 种基金Major Basic Research Project of Science and Technology of Yunnan (202001BC070001 to Y.C.C.)Natural Science Foundation of Yunnan Province (202102AA100053 to Y.C.C.)。
文摘Neural tube defects(NTDs)are severe congenital neurodevelopmental disorders arising from incomplete neural tube closure.Although folate supplementation has been shown to mitigate the incidence of NTDs,some cases,often attributable to genetic factors,remain unpreventable.The SHROOM3 gene has been implicated in NTD cases that are unresponsive to folate supplementation;at present,however,the underlying mechanism remains unclear.Neural tube morphogenesis is a complex process involving the folding of the planar epithelium of the neural plate.To determine the role of SHROOM3 in early developmental morphogenesis,we established a neuroepithelial organoid culture system derived from cynomolgus monkeys to closely mimic the in vivo neural plate phase.Loss of SHROOM3 resulted in shorter neuroepithelial cells and smaller nuclei.These morphological changes were attributed to the insufficient recruitment of cytoskeletal proteins,namely fibrous actin(F-actin),myosin II,and phospho-myosin light chain(PMLC),to the apical side of the neuroepithelial cells.Notably,these defects were not rescued by folate supplementation.RNA sequencing revealed that differentially expressed genes were enriched in biological processes associated with cellular and organ morphogenesis.In summary,we established an authentic in vitro system to study NTDs and identified a novel mechanism for NTDs that are unresponsive to folate supplementation.
基金supported by the National Natural Science Foundation of China(No.32271363 and No.82172831)the Natural Science Foundation of Chongqing(No.cstc2021jcyj-msxmX0642).
文摘Lung cancer is emerging as a common malignancy worldwide,with non-small cell lung cancer(NSCLC)accounting for approximately 85%of all cases.Two-dimensional(2D)in vitro cell line cultures and animal models are currently used to study NSCLC.However,2D cell cultures fail to replicate the medication response and neoplastic heterogeneity of parental tumors.Animal models are expensive and require lengthy modeling cycles.The generation of in vitro three-dimensional(3D)tissue cultures called organoids,which exhibit multicellular,anatomical,and functional properties of real organs,is now achievable owing to advancements in stem cell culturing.The genetic,proteomic,morphological,and pharmacological characteristics of tumors are largely preserved in tumor organoids grown in vitro.The design and physiology of human organs can be precisely reconstructed in tumor organoids,opening new possibilities for complementing the use of animal models and studying human diseases.This review summarizes the development of NSCLC organoids and their applications in basic research,drug testing,immunotherapy,and individualized treatments.
基金Supported by Chinese Medicine Service System and Capacity Building(Key Project with Chinese Medicine Characteristics and Advantages,Ruikang Hospital,2023)Guangxi Science and Technology Major Project during the 14th five-year Plan,No.Guike AA22096028.
文摘Gastric organoids are models created in the laboratory using stem cells and sophisticated three-dimensional cell culture techniques.These models have shown great promise in providing valuable insights into gastric physiology and advanced disease research.This review comprehensively summarizes and analyzes the research advances in culture methods and techniques for adult stem cells and induced pluripotent stem cell-derived organoids,and patient-derived organoids.The potential value of gastric organoids in studying the pathogenesis of stomach-related diseases and facilitating drug screening is initially discussed.The construction of gastric organoids involves several key steps,including cell extraction and culture,three-dimensional structure formation,and functional expression.Simulating the structure and function of the human stomach by disease modeling with gastric organoids provides a platform to study the mechanism of gastric cancer induction by Helicobacter pylori.In addition,in drug screening and development,gastric organoids can be used as a key tool to evaluate drug efficacy and toxicity in preclinical trials.They can also be used for precision medicine according to the specific conditions of patients with gastric cancer,to assess drug resistance,and to predict the possibility of adverse reactions.However,despite the impressive progress in the field of gastric organoids,there are still many unknowns that need to be addressed,especially in the field of regenerative medicine.Meanwhile,the reproducibility and consistency of organoid cultures are major challenges that must be overcome.These challenges have had a significant impact on the development of gastric organoids.Nonetheless,as technology continues to advance,we can foresee more comprehensive research in the construction of gastric organoids.Such research will provide better solutions for the treatment of stomach-related diseases and personalized medicine.
基金Supported by the Chongqing Natural Science Foundation Project,No.CSTB2022NSCQ-MSX0172.
文摘BACKGROUND Patient-derived organoids(PDOs)have been demonstrated to predict the response to drugs in multiple cancer types.However,it remains unclear about its application in cholangiocarcinoma.CASE SUMMARY A 59-year-old woman was admitted to the hospital due to upper abdominal pain for over 8 months.According to relevant examinations,she was diagnosed as perihilar cholangiocarcinoma(pCCA)with intrahepatic metastasis and perihilar lymphatic metastasis.After multidisciplinary team discussion,percutaneous transhepatic cholangiodrainage was performed to relieve biliary obstruction,and puncture biopsy was conducted to confirm the pathological diagnosis.Transarterial chemoembolization with nab-paclitaxel was used in combination with toripalimab and lenvatinib,but the levels of tumor markers including alpha fetal protein,carcinoembryonic antigen,carbohydrate antigen 15-3 and cancer antigen 125 were still raised.The PDO for drug screening showed sensitive to gemcitabine and cisplatin.Accordingly,the chemotherapy regimen was adjusted to gemcitabine and cisplatin in combination with toripalimab and lenvatinib.After 4 cycles of treatment,the tumor was assessed resectable,and radical surgical resection was performed successfully.One year after surgery,the patient was still alive,and no recurrence or occurred.CONCLUSION PDOs for drug sensitivity contribute to screening effective chemotherapy drugs for advanced pCCA,promoting conversion therapy and improving the prognosis.
