The olfactory mucosa serves as both a sensory organ and an immune barrier to protect against bacterial and viral invasion and other insults. It is unclear how different types of olfactory mucosal cells coordinate and contribute to these two functions. We set out to reveal the critical roles of a subset of microvillous cells of the mucosa, olfactory tuft cells, in protecting and reconstructing this vital olfactory sensory organ. We first validated the expression of canonical gustatory signalling proteins and other molecular markers in olfactory tuft cells. Genetic disruption of the Gng13 and Trpm5 genes that encode the two gustatory signalling proteins, G protein subunit Gγ13 and transient receptor potential ion channel Trpm5, respectively, resulted in elevated basal inflammation and enhanced activation of the quiescent stem cells-horizontal basal cells (HBCs) in the mucosa. Nasal infection of H1N1 influenza virus further exacerbated the inflammation and delayed the resolution of inflammation in the mutant mucosa, including more immune cell infiltration, augmented cytokine production and cell death, increased HBC proliferation and direct differentiation into tuft cells, and prolonged olfactory tuft cell hyperplasia. Cytokine treatment of the cultured olfactory epithelial organoids indicated that the cytokines that were found to be elevated in the mutant mucosa, including interleukin-4 (IL-4), IL-13 and interferon-γ (INF-γ), are able to stimulate HBC activation. Together, our results indicate that olfactory tuft cells play an important role in maintaining the baseline inflammation under the steady-state condition, and altering inflammatory magnitude and modulating HBC activation and differentiation in the olfactory mucosa following the viral infection. Our findings shed light on new roles of olfactory tuft cells in innate immune response, quiescent stem cell activation and neuroimmune interactions, and provide novel therapeutic targets for preventing and treating stem cell-related olfactory disorders such as chronic rhinosinusitis and long COVID.

Trophoblast stem cells (TSCs) have emerged as a valuable model for investigating placentation and related disorders. Despite their ability to be maintained over multiple passages without losing stemness, conventional TSCs are characterised by notorious heterogeneity and a spontaneous differentiation tendency. In this study, we established a type of mouse TSCs with dome-shaped colonies (dTSCs). The dTSCs expressed canonical markers of conventional TSCs and exhibited relatively low levels of heterogeneity and spontaneous differentiation when cultured in conventional trophoblast stem cell medium. Additionally, dTSCs retained robust potential to differentiate into all trophoblast lineages in vitro, as well as the capacity to contribute to placental development in vivo. Furthermore, the functions of Foxo4 and Slc16a3 (MCT4) could be effectively explored in trophoblast development by using our established dTSCs. Collectively, our findings establish a type of high-quality TSCs and define dTSCs as a promising and refined model for investigating trophoblast development.

Residual PSC detection follows ultra‑trace‑analysis principles. LOD supports qualitative assessment. Assays employ PSC‑specific markers and proper references, with adaptation to differentiated cell populations from diverse PSC sources.

This document specifies the technical elements for bacteria and fungi detection by PCR assay which enables rapid, broad-spectrum detection of bacteria and fungi with high sensitivity. Qualitative judgement is based on LOD. Key detection points include efficient nucleic acid extraction, broad-spectrum primer-probe design, high amplification efficiency and minimised background interference.

Polyploidy is normally found in numerous tissues and cell types in the generally diploid human body and is crucial for proper development. However, polyploidy, and especially triploidy, can also be found in malignant tumours and is often associated with chemoresistance, stemness and metastatic capabilities. Here, we utilise our isogenic haploid, diploid and triploid human embryonic stem cell (hESC) lines to study the effect of ploidy on the response to four different anticancer drugs. Surprisingly, we show that triploid cells are more sensitive to chemotherapy-induced apoptosis and cell cycle arrest than diploid and haploid cells, correlated with higher levels of DNA damage. This phenotype is regulated by p53, as it was reversed in TP53-KO cells, where triploid mutant cells display higher resistance to the chemotherapies we applied compared to mutant haploid and diploid cells. The reversal from sensitivity to resistance, driven by TP53-KO of the triploid cells, was accompanied by a reversal in the enrichment of DNA repair, replication and cell division related genes, compared to their diploid counterparts. Conversely, triploidy triggered DNA damage-induced differentiation in both WT and TP53-KO treated cells, pointing to a ploidy-dependent response not mediated by p53. In addition, we show that cancer cell lines also display similar ploidy-dependent anticancer drug responses to our TP53-KO hESCs. These findings uncover the interplay between ploidy and the p53 pathway in determining the outcome of anticancer therapeutics, and display the potential use of isogenic hESCs, differing only in their ploidy level, in studying the impact of ploidy on chemotherapy response.

Persistent inflammation, insufficient vascularization and impaired osteogenesis limit the repair of critical-sized femoral defects. Although microRNA-210-3p (miR-210-3p) can regulate these processes, its therapeutic use is constrained by poor extracellular stability and inefficient cellular entry. Here, miR-210-3p was incorporated into a tetrahedral framework nucleic acid (tFNA) to create tFNAs-miR-210-3p. Dynamic light scattering, zeta-potential analysis, atomic force microscopy (AFM) and capillary electrophoresis supported formation of the negatively charged nanosystem. In RAW264.7 macrophages, tFNA delivery increased Cy5-labelled miR-210-3p-positive cells from 1.24% for free miR-210-3p to 99.93%. Under lipopolysaccharide (LPS)-induced inflammatory conditions, tFNAs-miR-210-3p reduced CD86 and inducible nitric oxide synthase, increased CD206 and arginase-1, enhanced endothelial tube formation and improved osteogenic differentiation. It suppressed M1 marker CD86/iNOS and elevated M2 marker CD206/Arg-1 to resolve the inflammatory microenvironment. Mechanistically, miR-210-3p directly binds the 3'UTR of EFNA3 mRNA to inhibit its expression, thereby activating AKT/STAT3 signalling to switch macrophages toward reparative phenotype. In a rat critical-sized femoral defect model, local tFNAs-miR-210-3p treatment improved histological repair, macrophage polarisation, microcomputed-tomography indices, vascularization and osteocalcin expression at 4 and 8 weeks. These findings identify tFNA-mediated miR-210-3p delivery as a promising immunomodulatory strategy for vascularised bone regeneration.

Vascularised lung organoids (vLOs) that faithfully mimic human lung tissue architecture and disease pathology are critical for advancing pulmonary research but remain challenging to generate. Here, we developed a robust self-organisation protocol to produce vLOs with cellular heterogeneity and functional vasculature. The engineered blood vessels within vLOs exhibit lung-specific characteristics. As proof of concept, we applied this platform to model chronic obstructive pulmonary disease (COPD) and pulmonary hypertension (PH). Using patient-derived vLOs, we demonstrated that cigarette smoke extract (CSE) induces pathological features resembling clinical COPD, including epithelial disruption and inflammatory responses. Furthermore, vLOs generated from a PH patient recapitulated disease-associated vascular remodelling, with RNA-seq revealing dysregulated pathways in endothelial dysfunction. Notably, we identified sodium hydrosulfide (NaHS) as a potential therapeutic candidate, as it attenuated aberrant EndoMT in PH-vLOs. In summary, our study establishes a physiologically relevant vLO system that enables modelling of cell-type-specific disease mechanisms and underscores the broad utility of this platform for mechanistic investigations and precision medicine approaches in respiratory disorders.

CKIP-1, as a scaffold protein with special structural domains, treats related diseases through multiple emerging targeted technologies.

This guideline provides standardized requirements and quality control measures for human gallbladder cancer organoids, supporting their reliable application in research and precision medicine. It aims to promote methodological consistency, facilitate international collaboration, and accelerate translational research and clinical implementation.

To streamline the application of sequence-based DL methods in single-cell genomics, we established a two-layer CNN as our baseline model. We focus our benchmark on how data characteristics, hyperparameter optimization, and advanced model architectures impact performance across sequence-to-expression and sequence-to-regulation tasks. A key contribution of our study is the exploration of multi-task learning (MTL) frameworks for mitigate technical sparsity. We demonstrated that MTL significantly enhances the modeling of cellular heterogeneity, evaluating the effectiveness of task grouping and balancing strategies, with particular focus on the prediction of rare cell types. Our comprehensive comparative analysis provided an actionable framework and valuable insights for guiding future research endeavors and facilitating the development of the sequence-based DL models capable of superior predictive performance in single-cell genomics.

Intervertebral disc degeneration (IVDD) is a major contributor to low back pain, but the immune-metabolic events that sustain disc inflammation remain poorly defined. Here, we investigated whether disturbed macrophage lipid handling promotes IVDD by coupling cellular senescence to defective efferocytosis. Clinical magnetic resonance imaging and biochemical profiling showed that advanced IVDD was accompanied by increased disc fat fraction and systemic lipid abnormalities. Mendelian randomisation, bulk transcriptomics, machine-learning modelling and single-cell RNA sequencing further linked lipid metabolic disturbance to immune remodelling in degenerative discs, and identified TIAM2, SLC44A4, PPT1 and PTGDS as lipid metabolism-related hub genes associated with IVDD. At the single-cell level, disc macrophages with low glycerophospholipid metabolism scores showed higher senescence activity, a more inflammatory polarisation state and reduced efferocytosis-related signatures. Functional studies in bone marrow-derived macrophages indicated that lipid overload promoted lipid droplet accumulation, lipid peroxidation, impaired lipophagy and cholesterol efflux, activation of the p53/p21 senescence pathway, and reduced apoptotic nucleus pulposus cell clearance. Restoring lipid homeostasis with rosuvastatin enhanced lipophagy, recovered ABCA1/ABCG1-mediated cholesterol export, attenuated macrophage senescence and inflammatory activation and improved efferocytosis. In a rat needle puncture model, local rosuvastatin delivery alleviated disc structural damage, preserved proteoglycan content, balanced extracellular matrix metabolism and reduced macrophage senescence and inflammatory markers. In conclusion, this study supports macrophage lipid homeostasis as an important regulator of IVDD-associated immune dysfunction through senescence-associated impairment of efferocytosis.

Cystic fibrosis (CF) has shifted from a fatal paediatric lung disease to a multi-organ, lifespan-spanning disorder in which gastrointestinal (GI) complications and malignancies are increasingly prominent. With improved survival into mid- and late adulthood, aided by newborn screening, optimised nutrition and the advent of highly effective CF transmembrane conductance regulator (CFTR) modulators, implementation of colonoscopic surveillance has highlighted a several-fold increase in early-onset colorectal cancer (CRC) and a broader spectrum of intestinal pathology. In parallel, work in mouse models, human tissue and patient-derived intestinal organoids now places the CFTR at the centre of a complex epithelial compartment that integrates ion and pH homeostasis, mucus biology, microbiota, redox balance and immune/stromal function. In this review, we advance the hypothesis that CF may be conceptualised as a niche-centric hereditary CRC predisposition syndrome, in which germline CFTR dysfunction chronically destabilises epithelial identity in addition to increasing mutational burden. We synthesise evidence that CFTR loss remodels stem cell regulation, promotes hypoxia and oxidative stress, perturbs microbial ecosystems, drives chronic immune activation and stromal remodelling, and induces epithelial-mesenchymal plasticity (EMP) and DNA-damage vulnerability, collectively creating a pre-neoplastic intestinal ecosystem. We situate this model within contemporary CRC frameworks that emphasise cell-state transitions, hybrid epithelial-mesenchymal (E/M) states and specialised stromal niches, and we distinguish it from oncofoetal reprogramming, an APC-driven programme that CF does not clearly recapitulate. We propose that CF may provide a naturally occurring human context in which chronic epithelial stress sustains EMP-like pressure, a concept that requires direct validation in human CF intestinal tissue. Finally, we consider how CFTR modulators, microbiome-directed therapies and redox-targeted interventions might re-programme the CF intestinal niche and outline experimental and clinical strategies needed to determine whether early, ecosystem-level correction can prevent GI cancers in this high-risk population.

Liver organoids are three-dimensional miniature liver models that recapitulate the complex architecture and key functions of the human liver in vitro, offering powerful platforms for both fundamental research and translational applications. This review systematically summarises current fabrication strategies, disease-modelling utilities and regenerative potentials of liver organoids, alongside the major challenges and future directions. In recent years, the field has witnessed several breakthroughs. Through endothelial co-culture approaches, vascularised and metabolically zonated liver organoids have been successfully generated, achieving endothelial coverage exceeding 85%. Prime editing enables precise correction of pathogenic mutations in patient-derived organoids, with no off-target effects detected at the genome-wide level. In disease modelling, iPSC-derived liver organoids faithfully recapitulate the pathological progression of metabolic dysfunction-associated steatotic liver disease (MASLD) and verify the lipid-lowering efficacy of semaglutide. Macrophage-integrated organoid models support the full life cycles of HEV, SARS-CoV-2 and dengue virus, providing new tools for antiviral drug screening. Large-scale patient-derived tumour organoid biobanks successfully preserve the heterogeneity and clinical drug-resistance signatures of liver cancers. In regenerative medicine, encapsulated hepatocyte organoids and the UTOpiA bioartificial liver system have effectively rescued acute liver failure in animal models, while gene-edited autologous organoids offer potential curative strategies for genetic disorders such as Wilson disease. Nevertheless, insufficient hepatocyte functional maturity, difficulties in constructing vascular networks, and the lack of standardised culture protocols remain major obstacles to clinical translation. By bridging fundamental liver biology and clinical practice, liver organoid technology lays a solid foundation for precision hepatology and regenerative therapies. Continued interdisciplinary efforts are still required to overcome current limitations and facilitate its clinical adoption.

Dentinogenesis, a process essential for tooth function, relies on the precise differentiation of dental papilla cells into odontoblasts. This lineage commitment is governed by complex transcription factor networks. RUNX2 and KLF4, known key TFs co-expressed in this process, were proven to operate synergistically, but the specific cofactors coordinating their activity at the chromatin level remain unknown. Here, we identify the chromatin remodeller BRD9 as a novel interactor of both RUNX2 and KLF4 during odontoblastic differentiation. To probe their shared function, we generated neural crest-specific conditional knockout mice. Wnt1-Cre; Brd9[fl/fl] mice exhibited a disordered odontoblast layer with reduced secretion of extracellular matrix proteins (DMP1 and DSPP), strikingly phenocopying the compound Wnt1-Cre; Runx2[fl/wt]; Klf4[fl/fl] mutants. In vitro, chemical BRD9 degradation suppressed odontoblastic differentiation and mineral deposition. Integrative analyses of RNA-seq and ATAC-seq revealed that BRD9 maintained chromatin accessibility at odontogenesis-associated regions enriched with RUNX2 and KLF4 motifs, specifically at the Fam20c enhancer, thereby facilitating RUNX2 and KLF4 binding for transcriptional activation. Crucially, exogenous supplementation of FAM20C protein partially rescued the odontoblastic differentiation defects upon BRD9 loss. These findings establish BRD9 as a critical epigenetic orchestrator that coordinates RUNX2-KLF4 synergy to activate key odontogenic genes like Fam20c, thus affecting odontoblastic differentiation and dentinogenesis.