Search Results (871)

Three-dimensional (3D) cell culture models more faithfully reproduce native tissue organization and function than conventional two-dimensional systems, yet many existing bioprinting methods depend on scaffolds, complex instrumentation, or limited control over cell positioning. This review examines magnetic bioinks as a versatile platform for contactless 3D cell manipulation and biofabrication. It first outlines the fundamentals of magnetophoresis and defines magnetic bioinks as combinations of magnetic agents, including magnetic nanoparticles or paramagnetic salts, with biological components such as cells, proteins, or fluids. The review then compares label-based strategies, in which cells are magnetized and guided by positive magnetophoresis, with label-free approaches that exploit magnetic susceptibility differences to position diamagnetic cells through negative magnetophoresis. Across these methods, magnetic bioinks have enabled single-cell sorting, spatial patterning, spheroid and co-culture assembly, multilayer tissue formation, and hydrogel-integrated printing. These capabilities support applications in disease modeling, drug screening, biosensing, regenerative medicine, and emerging biofabrication under microgravity conditions. The paper also highlights key limitations, including nanoparticle biocompatibility, paramagnetic salt toxicity, osmotic stress, and the need for better assay standardization and translational validation. Overall, magnetic bioinks represent a promising scaffold-free approach for rapidly producing physiologically relevant 3D biological constructs for research and clinical innovation. Full article

Porous hydrogels are critical for tissue engineering and regenerative medicine, as they mimic the native extracellular matrix to support cell infiltration and mass transport. A common strategy for engineering pore structures involves the incorporation and subsequent removal of sacrificial porogen templates (e.g., crystals or microspheres). Although this approach offers excellent control over pore architecture, it often suffers from complex procedures and biosafety concerns arising from incomplete template removal. In this work, we present a simple, biocompatible, and versatile templating approach. By systematically investigating the coacervation parameters, we produced gelatin microspheres (GSs) with tunable diameters from 7 µm to 300 µm via a green, instrument-free, and scalable process. Using GSs of 20–160 µm as porogens, we obtained alginate hydrogels with adjustable viscoelasticity, stiffness, and pore sizes. We then validated two cell-loading strategies for bulk porous alginate hydrogels using immortalized human T (Jurkat) cells: (i) post-seeding into pre-formed pores supported high-density, long-term, and organized cell aggregates with >90% viability; (ii) in situ encapsulation (prior to pore formation) yielded >80% viability and preserved the cluster-forming growth characteristics of Jurkat cells. Moreover, composites of smaller GSs (7–20 µm) with alginate could be syringe-extruded into stable, sub-millimeter porous filaments, demonstrating the potential for 3D printing. Collectively, this work provides a promising platform for three-dimensional culture of immune cells. Full article

Background: Yes-associated protein 1 (YAP1), a key effector of the Hippo signaling pathway and mechanosensitive transcriptional coactivator, plays a complex role in osteoarthritis (OA) and cartilage regeneration. While YAP1 is essential for tissue homeostasis, its dysregulation has been implicated in both inflammatory and degenerative joint pathologies. However, its precise function remains ambiguous. Methods: We silenced YAP1 with small interfering RNA (siYAP1) in two human-cell-based models relevant to OA pathogenesis and cartilage repair: (1) IL-1β (10 ng/mL)-stimulated articular chondrocytes in monolayer and pellet cultures, and (2) TGF-β1 (10 ng/mL)-induced chondrogenesis in MSC pellet cultures. Outcome measures comprised YAP1 nuclear localization; inflammatory/catabolic markers in chondrocytes (IL6, IL8, ADAMTS5, MMP13); and, in MSC pellets, chondrogenic or hypertrophic markers (COL2A1, ACAN, RUNX2, MMP13, COL10A1) together with glycosaminoglycan (GAG) deposition. Statistical significance was assessed using an ANOVA or Friedman test with post hoc correction (Tukey or Dunn’s test, respectively); p < 0.05 was considered significant. Results: In human chondrocytes, siYAP1 reduced IL-1β-induced nuclear YAP1 localization and suppressed pro-inflammatory mediators IL6 and IL8, indicating an anti-inflammatory effect. YAP1 silencing also downregulated ADAMTS5 expression in 2D monolayers but not in 3D pellet cultures, suggesting reduced regulatory influence in the three-dimensional environment. Notably, MMP13 expression was paradoxically increased following YAP1 knockdown, underscoring the complexity of YAP1’s role in catabolic regulation. In MSC chondrogenesis, siYAP1 enhanced TGF-β1-induced chondrogenesis by increasing COL2A1 and ACAN expression and promoting GAG deposition on day 21. Additionally, it reduced hypertrophic markers RUNX2 and MMP13 on day 7, though COL10A1 remained elevated compared to negative siRNA, indicating only partial suppression of hypertrophic differentiation. Nuclear YAP1 levels were increased by day 21 despite reduced mRNA, suggesting post-transcriptional regulation or enhanced nuclear translocation. Conclusions: These findings demonstrate that YAP1 knockdown exerts context-specific anti-inflammatory and pro-chondrogenic effects while partially mitigating hypertrophy. However, divergent outcomes, namely elevated MMP13 in chondrocytes and upregulated COL10A1 in MSCs, indicate that YAP1 silencing does not uniformly suppress inflammation or hypertrophy. YAP1 represents a potential therapeutic target for OA, but its modulation requires careful consideration of cellular context, siRNA delivery method, and timing to optimize outcomes for cartilage repair and joint preservation. Full article

Organoids are three-dimensional culture systems that self-organize and partially recapitulate the architecture, cellular composition, and functional properties of native tissues. In pediatric congenital hepatobiliary diseases, persistent cholestasis, bile duct maldevelopment, epithelial injury, and progressive fibrosis often lead to cirrhosis, liver failure, or the necessity for liver transplantation. Compared with conventional two-dimensional cell culture and animal models, hepatobiliary organoids provide patient-derived, human-relevant platforms for modeling disease mechanisms, evaluating therapeutic responses, and exploring regenerative strategies. Unlike previous reviews that mainly discuss general organoid culture systems or broad liver disease modeling, this review is organized around clinically oriented translational endpoints, including mechanistic target discovery, prognostic stratification, therapeutic validation, and regenerative reconstruction. We further discuss current barriers to clinical translation, including reproducibility, scalability, vascularization, immune integration, manufacturing standardization, and patient-specific genetic, environmental, and dietary modifiers. By integrating disease-specific mechanisms with translational applications, this review provides a framework for understanding how organoid-based platforms may contribute to future diagnosis, risk assessment, therapeutic decision-making, and regenerative medicine in pediatric congenital hepatobiliary disorders. Full article

Tumor heterogeneity and microenvironmental complexity remain fundamental barriers to genomics-centered precision oncology, frequently causing discordance between molecular alterations and real-world therapeutic responses. Here, we reviewed patient-derived organoid (PDO) technologies as functional platforms that complement molecular profiling by directly investigating patient-specific sensitivity, resistance, and microenvironment dependent vulnerability. We first summarize why conventional preclinical systems, two-dimensional cell lines and patient-derived xenografts, are limited by reduced biological fidelity, impractical turnaround time, and scalability for clinical decision support. We then synthesized organoid-based evidence across three representative disease malignancies with distinct precision-medicine bottlenecks. Across these settings, we highlight advances that extend the PDO capability beyond the tumor epithelium alone, including air–liquid interface cultures, immune and stromal co-cultures, and microfluidic organoid-on-chip systems, as well as integration with multi-omics and artificial intelligence for scalable analytics. Finally, we discuss the key translational requirements, standardization of culture matrices and assay readouts, quality control, automation to reduce turnaround time, and regulatory/ethical frameworks, required to transition organoid-guided testing from proof-of-concept to routine implementation. Collectively, this review reframes organoids as functional stratification platforms supporting the integration of functional response profiling alongside genomics-guided precision oncology approaches. Full article

Nature-based tourism is expanding rapidly, placing new pressures on fragile ecosystems and governance structures that were not designed for the intensity and diversity of today’s visitors. Despite this trend, protected areas face unique management constraints and rapid socio-environmental changes. While motivational segmentation of tourists can provide valuable information to policymakers, this subject remains understudied/under-researched. This study addresses the gap by examining the motivations, behaviours, and attitudes of visitors to Romania’s national and natural parks, using a structured survey (n = 509) and a two-step approach combining dimensionality reduction with visitor segmentation. Principal component analysis (PCA) reveals distinct motivational dimensions related to visitors’ desire for immersion in nature, wildlife observation and learning, active recreation, and social–cultural engagement. Based on these dimensions, three visitor segments emerge through cluster analysis, with significantly different patterns of landscape use, expectations of recreational services, and perceptions of interpretation media. This research provides practical insights for targeted communication, zoning, and adaptive governance and proposes integrating visitor typologies with park management to support sustainable rural development. The findings highlight how a nuanced understanding of tourist segments can inform more effective policy measures that balance recreational demand with the long-term protection of natural and cultural resources, offering practical value for the sustainable development of protected areas, local communities, and other stakeholders. Full article

This systematic review examines how Extended Reality (XR) technologies, i.e., Virtual (VR), Augmented (AR), Mixed (MR), and Spatial Augmented Reality (SAR) are designed, implemented, and evaluated in cultural heritage (CH) applications, addressing five research questions: (RQ1) How were XR technologies applied in CH between 2021 and 2025? (RQ2) What interaction paradigms are used, and how do they shape engagement and meaning making? (RQ3) What user experience outcomes are reported in XR CH applications? (RQ4) What evaluation methods are employed and what methodological gaps remain? (RQ5) What challenges persist across XR heritage implementations? Peer-reviewed, English-language studies reporting on implemented XR systems in CH contexts with empirical or evaluative data were included; conceptual articles without a described implementation, non-English publications, and studies published before January 2020 were excluded. Scopus, Web of Science, IEEE Xplore, and the ACM Digital Library were searched for publications dated January 2020 through March 2025, complemented by manual proceedings screening (SIGGRAPH, CHI, IMX, VRCAI) and backward/forward citation tracking. All databases were last searched in March 2025. Two independent researchers screened all records and extracted data; disagreements were resolved through structured discussion. Bias toward positive novelty outcomes was mitigated by including conference proceedings alongside journal articles to broaden the evidence base. A qualitative thematic synthesis was employed, as methodological heterogeneity across studies precluded statistical meta-analysis. Findings were organized inductively into four thematic domains through iterative coding and inter-author consensus. From an initial corpus of 359 records, 287 unique records were retained after deduplication; following title/abstract screening and full-text eligibility assessment, 64 studies were included in the final synthesis. The majority (60/64) were published between 2021 and 2025, with study sample sizes ranging from small expert cohorts (n ≈ 6) to large public deployments (n > 125). The thematic analysis across technology, interaction design, user experience, and evaluation reveals trends toward participatory, multiuser, and multimodal XR designs, reporting benefits including immersion, engagement, learning, and accessibility, alongside recurring challenges such as cost, usability, cybersickness, content authenticity, and lack of longitudinal evaluation. Beyond thematic description, using a cross-domain analytical synthesis, we identify the Design Coherence Framework for XR Heritage (DCF-XR); this is a four-dimensional interpretive model spanning technology, interaction design, user experience, and evaluation, which provides an original diagnostic lens for understanding the conditions under which XR effectively serves cultural heritage goals. A typology of four recurring design failure modes, derived inductively from the corpus, demonstrates that the most persistent shortcomings in the field arise not from the weakness of individual dimensions but from their misalignment with one another. Evidence is limited by the predominance of small convenience samples, single-session laboratory evaluations, and the absence of domain-specific standardized assessment instruments for XR in CH, which constrains the generalizability of reported outcomes. Targeted recommendations for rigorous, ethical, and inclusive XR practice in CH are presented, highlighting the need for longitudinal studies, open datasets, and standardized evaluation frameworks. This review received no external funding. This review was not pre-registered in a prospective register. Full article

This Perspective is dedicated to the memory of Professor Joseph Mark (Joe) Gani (1924–2016) and Professor Christopher Charles (Chris) Heyde (1939–2008), two scholars whose intellectual leadership profoundly shaped applied probability, mathematical statistics, and their interface with finance, insurance, and risk management. Their contributions extend beyond specific technical results to the development of research cultures grounded in probabilistic rigor, empirical relevance, and methodological transparency. We emphasize three enduring themes central to modern quantitative risk analysis. First, the systematic incorporation of heavy-tailed and non-Gaussian features in stochastic modeling, reflecting persistent empirical deviations from classical Gaussian assumptions in financial data. Second, the development of stochastic and time-series methodologies capable of handling dependence structures, including conditional heteroskedasticity and long-range dependence. Third, the principled integration of probabilistic modeling with data-driven and machine learning approaches, ensuring predictive performance is accompanied by interpretability and robustness. We situate these contributions within contemporary challenges in financial risk management, including systemic risk, environmental, social and governance (ESG) considerations, and climate finance. In particular, climate-related financial risks arise from both physical impacts (such as extreme weather events and long-term environmental change) and transition dynamics associated with the shift toward a low-carbon economy (including policy, technological, and market adjustments). These sources of risk introduce additional forms of dependence, nonlinearity, and model uncertainty, particularly in high-dimensional, data-rich settings. This Perspective highlights a forward-looking research agenda that preserves the foundational principles of applied probability while adapting them to modern financial systems characterized by real-time information flows and evolving risk structures. This legacy continues to shape how financial risk is modeled, measured, and understood in increasingly complex and interconnected environments. Full article

Glioblastoma multiforme (GBM) remains the most aggressive and therapeutically intractable primary brain tumor, with many patients experiencing rapid relapse despite maximal surgical resection followed by standard chemoradiation. This persistent failure reflects the convergence of profound tumor-intrinsic genetic heterogeneity and a highly dynamic, spatially structured, and immunosuppressive tumor microenvironment (TME). Together, these forces create strong selective pressures that fuel tumor evolution, intratumoral diversity, phenotype plasticity, diffuse invasion, and robust resistance to therapy. The TME of GBM is orchestrated through a complex interplay between diverse cellular constituents, including tumor-associated macrophages, reactive astrocytes, endothelial cells, pericytes, and GBM stem cells, and non-cellular components such as extracellular matrix remodeling, hypoxia, metabolic and nutrient gradients, and spatially patterned cytokine and chemokine signaling networks. Additionally, heterogeneity in blood–brain barrier (BBB) and blood–tumor barrier (BTB) complicates drug delivery and immune surveillance, reinforcing therapeutic resistance and regional tumor adaptation. Conventional two-dimensional cell cultures and animal models fail to sufficiently capture these multiscale, patient-specific interactions, limiting their translational predictive power. In this narrative review, we synthesize recent advances in GBM organoid technologies as physiologically relevant, three-dimensional platforms that more faithfully recapitulate TME for driving tumor evolution and treatment resistance. We compare complementary organoid strategies, including patient-derived GBM organoids that preserve native cytoarchitecture, cerebral organoid co-culture systems that reconstruct tumor–brain interactions, and advanced platforms incorporating immune and vascular features such as air–liquid interface cultures, microglia-enriched systems, and BBB/BTB-integrated models. Finally, we highlight emerging innovations such as spatial transcriptomics, organoid-on-a-chip systems, live imaging coupled with lineage tracing, genome engineering, and artificial intelligence integration that collectively position GBM organoids at the forefront of precision neuro-oncology, reproducing TME, enabling dynamic mapping of tumor evolution, and accelerating patient-specific therapeutic discovery. Full article

Metabolic dysfunction-associated steatotic liver disease (MASLD) is a widespread condition with a complex pathogenesis. Cell-based models are important tools for studying the mechanisms underlying its development and progression. The aim of this review is to analyze the HepaRG, Huh-7, immortalized human hepatocyte (IHH), and primary human hepatocyte (PHH) cell lines for modeling and studying MASLD. HepaRG represents the most metabolically competent immortalized hepatocyte model with preserved biotransformation activity and a physiological bioenergetic response to lipid loading, making it valuable for pharmacological and toxicological studies. Huh-7 is distinguished by its accessibility and suitability for studying steatosis, lipotoxicity, insulin resistance, and paracrine mechanisms of fibrogenesis; however, its use is limited by its tumor origin, impaired carbohydrate metabolism, and low activity of xenobiotic-metabolizing enzymes. The IHH model occupies an intermediate position because of its non-tumor origin and is of interest for studies of senescence, epigenetic regulation, and signaling pathways involved in steatosis, although interpretation of results requires consideration of immortalization-related effects and specific metabolic limitations. PHH remains the most physiologically relevant platform for MASLD modeling, particularly in three-dimensional (3D) and microphysiological formats; however, its use is limited by high cost, interindividual variability, and the limited duration of the differentiated phenotype. Increasing model complexity—from two-dimensional (2D) monocultures to co-cultures, spheroids, and organ-on-chip systems—enhances physiological relevance and enables reproduction not only of steatosis but also of the inflammatory and fibrogenic components of MASLD progression, yet it reduces reproducibility and complicates standardization. Overall, none of the existing models is universal, and the optimal strategy is to select models according to the specific research question. A key direction for future research is the standardization of steatosis induction protocols and the unification of criteria for evaluating results. Full article

Traditional urban landscape evaluations have primarily relied on either objective spatial metrics, such as the Green View Index (GVI), or subjective human surveys, often failing to capture the complex mechanisms of human environmental perception. This study proposes a novel Explainable Artificial Intelligence (XAI) framework that integrates objective physical configuration with subjective cognitive assessment to predict human landscape preference. Utilizing 159 urban landscape images, we extracted physical features via semantic segmentation (SegFormer) and psychological perceptions via a zero-shot vision-language model (CLIP). Our hybrid Random Forest model successfully bridged these dimensions, achieving moderate yet promising predictive performance (Rsquare = 0.442). SHAP (Shapley Additive exPlanations) analysis revealed that psychological perceptions—specifically Safety (0.104), Fascination (0.096), and Tranquility (0.080)—outperformed traditional objective metrics like GVI (0.067) in determining overall preference, while sub-model interpretation linked these psychological responses to specific physical elements such as buildings, sky openness, low vegetation, and water bodies. The findings suggest that urban green space design should move beyond maximizing greenery quantity and instead prioritize spatial compositions that induce psychological security, visual interest, and restoration. The proposed framework offers a scalable and interpretable tool for human-centered landscape assessment, while acknowledging limitations related to sample size, cultural generalizability, pretrained model bias, and reliance on static two-dimensional imagery. Full article

Background: Impaired angiogenesis and persistent inflammation are hallmarks of chronic diabetic wounds. Extracellular vesicles derived from dental pulp stem cells (DPSC-EVs) represent a promising cell-free therapy for tissue repair; however, their clinical translation is hindered by suboptimal yields and attenuated bioactivity associated with conventional two-dimensional (2D) culture. This study investigated whether a biomimetic three-dimensional (3D) fibrin/gelatin hydrogel system could optimize the therapeutic potency of DPSC-EVs for diabetic wound healing. Methods: DPSCs were encapsulated within 3D fibrin/gelatin scaffolds, followed by comprehensive characterization of cell viability and morphology. 3D-EVs and 2D-EVs were isolated via ultracentrifugation and validated by transmission electron microscopy and nanoparticle tracking analysis. The pro-angiogenic capacity of 3D-EVs was evaluated using human umbilical vein endothelial cells (HUVECs) under high-glucose (HG) stress. Additionally, the immunomodulatory effects were assessed by monitoring macrophage polarization in lipopolysaccharide-stimulated RAW 264.7 cells. The therapeutic efficacy was further validated in vivo using a streptozotocin (STZ)-induced diabetic mouse model with full-thickness cutaneous wounds. Results: The 3D fibrin/gelatin hydrogel provided a supportive microenvironment that significantly augmented the secretory productivity of DPSCs. Compared to 2D-EVs, 3D-EVs exhibited superior functional resilience in restoring HUVEC migration and tube formation under HG-induced oxidative stress. Furthermore, 3D-EVs effectively orchestrated the macrophage transition from a pro-inflammatory M1 phenotype toward an anti-inflammatory M2 phenotype, thereby modulating the immune microenvironment. In vivo, topical administration of 3D-EVs markedly accelerated wound closure, promoted re-epithelialization, and enhanced microvascular density and collagen maturation in diabetic mice. Conclusions: Our findings demonstrate that the 3D fibrin/gelatin culture system effectively primes the therapeutic profile of DPSC-EVs. These engineered vesicles accelerate diabetic wound healing by synergistically promoting angiogenesis and resolving chronic inflammation, offering a robust and potent cell-free strategy for the management of chronic diabetic ulcers. Full article

In the digital economy era, Urban vitality has transitioned into an intertwined Virtual-Physical system. This study examines Changsha’s five urban districts through a dual-dimensional framework bridging physical (social, economic, cultural, and ecological) and virtual (video, social, and digital life) dimensions. Integrating Coupling Coordination Degree (CCD) and XGBoost-SHAP models, we elucidate the spatial patterns and nonlinear drivers of Virtual-Physical synergy. The results indicate that: (1) Urban Vitality exhibits a significant center-periphery gradient. Although the Coupling Degree between the two dimensions is high, the overall CCD remains relatively low, reflecting pervasive spatial mismatches. Notably, 55 units display a reverse pattern where Virtual Vitality surpasses Physical Vitality, suggesting that digital flows can reconfigure urban space by transcending traditional locational constraints. (2) Interactions within the built environment exert pronounced threshold effects. Structural elements require specific critical masses to activate synergy, beyond which marginal returns diminish, as exemplified by the U-shaped effect of the Green View Index and the inverted U-shaped effect of Spatial Enclosure on CCD. (3) Interaction analysis identifies building density as a multiplier, unlocking the synergistic potential of land-use mix and transport networks once critical thresholds are surpassed. Furthermore, the efficacy of population and transit relies on dense road networks and intersection, while functional diversity buffers against negative micro-environmental impacts. This study advocates for a shift from facility-increment to threshold-triggered precision strategies in urban regeneration, providing empirical support for human-centric planning in the digital twin era. Full article

Owing to the multifactorial nature of inflammatory bowel disease (IBD) pathogenesis, conventional two-dimensional (2D) models inadequately recapitulate the complex in vivo microenvironment. This study sought to develop an immune-microenvironment-integrated intestinal-on-a-chip model to overcome these limitations. A microfluidic chip was engineered to co-culture intestinal epithelial (Caco-2) cells and macrophages, facilitating the simulation of IBD pathological conditions for mechanistic investigations. Following inflammatory stimulation, M0 macrophages polarized into the M1 phenotype, concomitant with the upregulation of pro-inflammatory cytokines, including tumor necrosis factor-alpha (TNF-α), interleukin-6 (IL-6), and interleukin-1 beta (IL-1β). This induction disrupted the expression of tight junction proteins (e.g., zonula occludens-1 [ZO-1]) in Caco-2 cells, thereby compromising epithelial barrier integrity. Infliximab was used as a model drug to inhibit TNF-α and modulate macrophage polarization within the chip, effectively rescuing impaired epithelial barrier integrity. This study establishes a reliable intestinal-on-a-chip model that recapitulates macrophage–epithelial interactions in IBD, providing a robust platform for elucidating the mechanisms underlying intestinal barrier dysfunction and developing targeted therapeutic strategies. Full article

Background: Corneal organoids derived from pluripotent stem cells have emerged as powerful tools for studying corneal development, disease modeling, and regenerative medicine applications. While previous protocols have successfully generated corneal tissue structures, there remains a need for three-dimensional models that recapitulate the complex cellular architecture and diversity of native human cornea. Methods: We developed a modified spontaneous three-dimensional corneal organoid model using human embryonic stem cells (hESCs) through an adapted Self-formed Ectoderm Autonomous Multi-zone (SEAM) protocol. hESCs were cultured as spheroids in ultra-low-binding plates under normoxic conditions and differentiated over 7–8 weeks. Organoids were characterized using immunofluorescence staining for corneal-specific markers and single-cell RNA sequencing to assess cellular composition and gene expression patterns. Results: Approximately 20% of organoids developed transparent regions characteristic of corneal tissue by day 30 of differentiation. Immunofluorescence analysis revealed spatially organized expression of corneal markers, including ZO-1 and E-cadherin in the outermost epithelial layers, P63α-positive putative limbal stem cells at the epithelial–stromal interface, vimentin-positive stromal cells in the interior, and laminin-1 deposition that suggests Bowman’s membrane formation. The organoids expressed cornea-specific keratins (K3, K12, and K15) and the master regulator PAX6 in appropriate cellular compartments. Single-cell RNA sequencing identified 18 distinct cell clusters, including three corneal epithelium subclusters with differential expression of MUC16, KRT12, and ΔNp63α, two stromal populations with distinct inflammatory profiles, and a corneal endothelium cluster. Transcriptomic analysis confirmed expression of key corneal genes, including AQP3, CDH1, multiple keratins, mucins, and extracellular matrix components (HAS2, CD34, CD44, COL8A1, and KERA). Conclusions: This three-dimensional spheroid-based putative corneal organoid model successfully recapitulates the multilayered architecture and cellular diversity of human cornea, including stratified epithelium, putative limbal stem cells, stroma, and endothelium in spatially appropriate arrangements. The model demonstrates molecular signatures consistent with native corneal tissue and provides a valuable platform for studying corneal development, disease mechanisms, and potential therapeutic applications. Future optimization to improve organoid formation efficiency and functional maturation will enhance the utility of this system for both basic research and translational medicine. Full article