Search Results (3,049)

The growing global demand for ethical, resource-efficient protein sources has renewed serious interest in cultured meat as a viable alternative to conventional livestock production. Two revolutionary biotechnological systems, induced pluripotent stem cell (iPSC) reprogramming and CRISPR-Cas9-mediated genome editing, when combined, offer unparalleled accuracy and scalability for the biofabrication of avian flesh. In this review, we present a comprehensive pipeline that involves the ectopic expression of Yamanaka factors (Oct4, Sox2, Klf4, and c-Myc) to reprogram primary somatic cells derived from Gallus gallus into induced pluripotent stem cells (iPSCs). This process is subsequently followed by targeted genome editing to enhance myogenic potential, growth efficiency, nutritional composition, and disease resistance. iPSCs are cultivated in a xeno-free bioreactor following genome editing, and subsequently directed to develop into myoblasts and mature myotubes. Three-dimensional tissue biofabrication is realized by combining biomaterial scaffolds and perfusion bioreactor systems, structuring an authentic muscle tissue matrix. These engineering platforms enable precise control over microenvironmental parameters, including oxygenation and nutrient perfusion. The resulting biofabricated poultry product is compositionally optimized, free of antibiotic residues, and exhibits a significantly reduced environmental footprint than poultry that is grown in the traditional way. This all-in-one solution solves important problems in food security, animal welfare, land use efficiency, and greenhouse gas emissions while also setting up a scalable biomanufacturing framework for making proteins for the next generation. Full article

Alzheimer’s Disease (AD) is a multifaceted progressive neurodegenerative disease characterized by memory deficits and cognitive impairment. The disease is clinically diagnosed by the presence of β-amyloid (Aβ), hyperphosphorylated tau, and neurodegeneration. Animal models serve as an indispensable tool to understanding AD pathogenesis and evaluating potential therapeutic approaches. However, despite the development of more than 200 rodent models, species-specific differences limit the translational relevance. Brain organoids generated from induced pluripotent stem cells (iPSCs) have the potential to bridge the gap between transgenic mouse models and clinical trials in human patients. Herein, we describe a robust stimulus-based neuroimmune organoid model that demonstrates neuroinflammation, neurodegeneration, and lipid dysregulation with AD-relevant pathological markers. Using planar organoids containing neurons, astrocytes, microglia, and vascular cells, we performed in-depth characterization of the induced AD-like phenotype using supernatant proteomic analysis, immunofluorescence staining, NfL and GFAP release, scRNAseq, bulk RNAseq, and pathway analysis both acutely (24 h) and chronically (7 days). Furthermore, we show that the induced neuroinflammation, lipid dysregulation, and neurodegeneration can be ameliorated using small molecules. This defined inducible model system presents an opportunity for drug discovery and development using a complex multicellular brain microenvironment derived from human iPSCs. Full article

Stage-specific embryonic antigen-3-positive pluripotent-like/macrophage-like multilineage-differentiating stress-enduring (Muse) cells are a distinct subpopulation of mesenchymal stromal cells (MSCs), accounting for 1% to several percent of MSCs. Although stem cell therapy for spinal cord injury (SCI) typically targets the subacute phase to avoid the hostile acute environment, the therapeutic window for Muse cells remains unclear. C57BL/6J mice with severe T9 contusion SCI received a single tail vein injection of human bone marrow-derived (BM) Muse cells, BM-MSCs (both 5 × 10⁴ cells), or vehicle at 2, 8, 14, or 28 days post-injury (DPI) without immunosuppressants. Among the different administration time points, the 2-DPI Muse cell group exhibited significantly higher Basso Mouse Scale scores than the BM-MSC and vehicle groups from 14 days after injection, while no significant differences were observed at the other administration time points. The 2-DPI Muse cell group showed significantly greater homing to the injured spinal cord than the BM-MSC group, with persistent engraftment and neural-lineage marker expression at day 42. Ablation of engrafted Muse cells at day 42 partially reversed locomotor recovery, suggesting that engrafted Muse cells contributed to functional recovery. These findings suggest that intravenous Muse cell therapy exerts timing-dependent therapeutic effects after SCI, with greater efficacy during the early post-injury phase. Full article

Several protocols have been reported for inducing parathyroid differentiation from human pluripotent stem cells in vitro. However, the efficiency of terminal differentiation into mature parathyroid cells remains limited. In this study, we performed comparative transcriptomic analyses of parathyroid lineage-induced human induced pluripotent stem (iPS) cells to clarify their developmental stage identity. Comparative analyses with adult parathyroid adenoma, used as a surrogate reference for mature parathyroid cells, and thymuses revealed that the differentiated cells exhibited gene expression patterns consistent with early-stage parathyroid and thymus development, including TBX1, which is required for fate determination of the parathyroid–thymic primordium. On the other hand, the expression of mature parathyroid marker genes, such as PTH, GCM2, and the calcium-sensing receptor (CaSR), remained markedly lower than that in parathyroid adenoma. Under the differentiation conditions and durations examined, application of modified differentiation protocols did not significantly enhance the expression of mature parathyroid markers, although pharyngeal pouch developmental gene expression was maintained. Our findings indicate that current in vitro differentiation systems effectively induce the parathyroid–thymic primordium stage, while a developmental bottleneck at the transition from the primordium toward parathyroid lineage commitment remains. Full article

Transparent conductive interfaces can enable optical pre-assessment of cardiac cell layers while remaining compatible with label-free electrophysiological recording. Here, we evaluated the integration of a monolayer graphene electrode into a capacitive recording platform for the analysis of human-induced pluripotent stem cell-derived cardiomyocyte (hiPSC-CM) monolayers. hiPSC-CMs cultured on graphene sensors formed confluent, synchronously beating monolayers that could be assessed by light microscopy prior to recording. Capacitive current transients could be recorded from spontaneously beating hiPSC-CM monolayers, supporting the compatibility of transparent graphene interfaces with capacitive recordings from electrically active cardiac cell layers. Signal amplitude and waveform morphology varied across sensors, indicating that recording performance depended strongly on the cell–sensor interface, including cell attachment, monolayer integrity, and capacitive coupling at the sensing surface. A descriptive perturbation sequence using the hERG blocker dofetilide revealed changes in waveform morphology and beat timing across sequential recordings. However, the data do not allow firm attribution to a compound-specific effect and are not intended for quantitative pharmacological characterization. Overall, the results support graphene as a transparent conductive cell–sensor interface, which should be interpreted in the context of cell–substrate interactions at the sensing surface. Combining optical pre-assessment with functional capacitive readout may support integrated workflows. Further studies will be needed to differentiate material-, interface-, and recording-related contributions and to establish reliable conditions for reproducible and scalable recordings. Full article

Background: A widely cited 2024 analysis reported that only 5% of therapeutic interventions tested in animals obtain regulatory approval for human use, reviving the debate about the predictive value of animal models. This review asks how the scientific community has interpreted that finding and what collective position is emerging. Methods: The literature citing the original study was assembled from public citation indexes and classified by argumentative stance into five categories: defences of animal experimentation, critiques, methodological and disease-specific analyses, the author’s own position, and work developing non-animal methods; no new animal data were generated. Results: Across the corpus, attempts to strengthen the validity of animal studies tended to reduce rather than reinforce their apparent translational signal; the low translation rate recurred across unrelated disease domains; and even defenders of animal models increasingly restricted their claims to specific contexts while conceding poor translation. In parallel, non-animal methods—including organoids, organ-on-chip systems, and computational models—were repeatedly described as equalling or surpassing animal models in several fields. Conclusions: The scientific, ethical, economic and regulatory cases now converge, supporting a deliberately planned transition toward human-relevant methods rather than its deferral. Full article

Primordial germ cells (PGCs) are the earliest precursors of gametes and essential cellular materials for poultry germplasm conservation and genetic modification. In this study, PGCs isolated from 2.5-day male and female Silkie chicken embryos were defined as circulating PGCs (cPGC_M and cPGC_F), whereas PGCs isolated from 8.5-day male and female embryos were defined as gonad-derived PGCs (gPGC_M and gPGC_F). RNA sequencing with three biological replicates per group was performed to characterize developmental-stage- and sex-associated transcriptional programs. Comparative transcriptomic analyses identified stage-associated differences in pathways related to cell–matrix interaction, focal adhesion, PI3K–Akt signaling, and stem cell pluripotency, with more differentially expressed genes detected in the female than in the male stage comparison. Sex-biased expression was detectable at the circulating stage, mainly reflecting W- and Z-linked expression differences, and the number of sex-biased genes was greater after gonadal colonization, primarily because more autosomal differentially expressed genes were detected. Candidate genes and pathways, including HINTW, DMRT1, SOX2, EDNRB, LPAR4, GFRA3, ECM–receptor interaction, ribosome-related modules, and calcium signaling, were associated with these comparisons and are presented as targets for future validation rather than as confirmed regulators. Because the study used three biological replicates per group, RT-qPCR corroboration was limited to six female stage-associated genes, and no functional perturbation assays were performed, the findings are descriptive and do not establish causal mechanisms of PGC migration, gonadal colonization, or sex differentiation. Within these limitations, the dataset provides an exploratory resource for subsequent studies of chicken PGC biology and biotechnology. Full article

Tau protein aggregation and spreading are central features of neurodegenerative diseases such as Alzheimer’s disease and frontotemporal dementia. Here, we investigated the role of phosphatidylinositol 4-kinase type IIIα (PI4KIIIα) in regulating tau propagation. We first used tau biosensor cells to demonstrate that both pharmacological inhibition and genetic reduction in PI4KIIIα effectively reduce the seeding of tau aggregation by extracellular seeds. To extend these findings to a more physiologically relevant system, we generated induced pluripotent stem (iPS) cell-derived cortical neurons carrying pathogenic MAPT mutations. These neurons rapidly acquired tauopathy-associated features, including expression of disease-relevant isoforms such as 4R tau, thereby enabling in vitro modeling of tau pathology. Using this model, we established phenotypic assays to monitor tau propagation and aggregation and applied them to test candidate small molecules. Notably, inhibition of PI4KIIIα consistently reduced seeding of tau assemblies in human neurons, highlighting this kinase as an important player in the seeding of tau pathology. Collectively, our work identifies PI4KIIIα as a regulator of tau pathology and provides new experimental platforms to dissect the molecular mechanisms of tau propagation. These findings open potential avenues for the development of strategies to slow or prevent tau-mediated neurodegeneration in the central nervous system. Full article

Rare genetic diseases are heterogeneous across mechanisms, trajectories, and treatment responses. To date, approved therapies remain available for only a small proportion of rare genetic diseases. Oligonucleotide-based RNA therapeutics, particularly antisense oligonucleotides (ASOs) and small interfering RNAs (siRNAs), offer a promising therapeutic avenue for rare genetic diseases with sequence-level precision. However, traditional preclinical paths may mis-predict human outcomes when disease biology diverges from animal models. New approach methodologies (NAMs), including patient-derived induced pluripotent stem cells (iPSCs), organoid models, and clinical-trials-in-a-dish (CTiD), aim to bring human biology earlier into the translational pipeline. NAMs enable variant-to-function studies, efficacy screening, and safety triage at clinically relevant speed and scale. While critics argue that NAMs are unvalidated and cannot replace preclinical animal models, proponents report that they are increasingly able to recapitulate human phenotypes and predict clinical liabilities, although their predictive validity remains context-dependent. Here, a front-loaded human filter refers to the use of human-derived systems early in development to support mechanistic interpretation, candidate prioritization, and early liability assessment before broader nonclinical evaluation. Recent studies pairing NAMs with ASOs support rapid, patient-specific preclinical screening in selected settings, while also showing the need for broader evidence on delivery, pharmacology, safety, and clinical relevance. This review places these developments within the translational realities of oligonucleotide-based therapeutics, including model fidelity, ASO chemistry and optimization, delivery challenges, pharmacology, regulatory pathways for individualized ASOs, and accessibility. We also propose a pragmatic validation framework to assess the scientific and translational credibility of NAMs across rare genetic diseases. Full article

The convergence of artificial intelligence (AI) and organoid technology represents a transformative advance toward precision and predictive medicine. Organoids derived from pluripotent stem cells or patient tissues provide physiologically relevant three-dimensional models that recapitulate key aspects of native organ architecture and function. However, intrinsic biological heterogeneity, high-content imaging outputs, and dynamic spatiotemporal processes pose significant analytical challenges that exceed the capacity of conventional approaches. Recent advances in AI and machine learning enable automated image segmentation, quantitative morphometric profiling, and predictive modeling of organoid growth, differentiation, and therapeutic response, thereby enhancing reproducibility and translational relevance. The integration of multimodal datasets, including imaging, genomics, transcriptomics, epigenomics, proteomics, and metabolomics, has further enabled the development of organoid-based digital twins and in silico disease simulations to optimize personalized therapy. AI-enabled organoid-on-a-chip platforms, cloud-based analytics, and federated learning frameworks are accelerating the emergence of scalable, privacy-preserving, and data-driven biomedical ecosystems. Despite these advances, critical challenges persist, including data standardization, model interpretability, ethical governance, and clinical validation. In contrast to existing reviews that emphasize isolated AI applications, this study proposes a unified translational framework integrating AI-driven image analytics, multi-omics integration, digital twins, and organoid-on-a-chip systems within a precision medicine paradigm. By synthesizing current developments, methodological advances, and emerging trends, this study highlights how AI-powered organoid platforms can bridge experimental biology and clinical decision-making, with broad implications for drug discovery, disease modeling, and regenerative medicine. This review aims to provide a comprehensive overview of artificial intelligence–enabled organoid platforms by integrating advances in image analytics, multi-omics data integration, digital twins, and microphysiological systems, while highlighting their potential applications and future directions in precision medicine, drug discovery, and regenerative healthcare. Full article

The 3D brain organoids have been widely used as a tool to study human brain development and disorders. Although angiogenesis and blood vascular endothelial cells play important roles in brain development and pathogenesis in neurological disorders, most 3D brain organoids lack inherent endothelial cells and need either the addition of differentiated endothelial cells or to be transplanted to animals to reconstitute such vascular structures. However, these approaches could miss the developmental interactions between angiogenesis and neurogenesis in the human brain. To reconstitute a 3D organoid mimicking the in vivo development of neural and vascular endothelial cells, we cultured iPSC-derived embryoid bodies and sequentially applied endothelial and neuronal induction media along with Matrigel embedding. The resulting 3D organoid consists of both neural cells and endothelial cells with vascular-like structures, as determined by immunostaining. With scRNA-Seq analysis, the organoid was confirmed to contain neural cell types similar to human brains, including a variety of excitatory and inhibitory neurons and glia. Furthermore, when compared with conventionally generated cerebral organoids without endothelial cells using RNA-Seq analysis, the vascular endothelial-containing neural organoids (EC-neural organoids) showed different gene profiles and favored angiogenesis and vasculogenesis. Of the differentially expressed genes, KRBA2 expression was found to be higher in neural cells and its inhibition by siRNA treatment resulted in decreased transcriptions of a variety of genes specific to neuronal differentiation but not genes specific to pluripotent stem cells such as OCT4. The EC-neural organoids also expressed receptors to SARS-CoV-2 at levels similar to human brains. This 3D EC-neural model provides a useful tool to study the interactions between vascular endothelial cells and neural cells in brain development and potentially for the study of neural infectious disorders where vascular endothelial cells are targets for infection and mediators for neural damage. Full article

Little is known about how nicotinic signalling in human astrocytes may contribute to the functional neurotoxicity of compounds related to tobacco alkaloids and neonicotinoid pesticides. We generated a single-cell Ca²⁺-imaging assay in induced pluripotent stem cell (iPSC)-derived astrocytes, and profiled functional expressions of some neurotoxicologically relevant receptors. Responses to pharmacological tool compounds indicated the expression of nicotinic, muscarinic, purinergic, glutamatergic receptors and voltage-gated Na⁺/Ca²⁺ channels. Closer investigation of the nicotinic system, e.g., using the alpha7 nicotinic acetylcholine receptor (nAChR)-selective positive allosteric modulator PNU-120596 and alpha7-preferring agonist (AR-R17779) demonstrated that Ca²⁺ signals elicited by nicotine and neonicotinoids are dominated by alpha7 nAChRs and depend on the downstream activation of L-type Ca²⁺ channels and tetrodotoxin-sensitive Na⁺ channels. Crosstalk of nAChR activation/desensitization was not observed for the inflammatory response elicited by TNF or for activation of glutamatergic or purinergic signalling. However, pre-stimulation of nAChR by neonicotinoids significantly blunted the response to the neurotransmitter acetylcholine. Comparative experiments in the human neuronal cultures (LUHMES cells) revealed similar potency ranges and pharmacological fingerprints for several neonicotinoids and their human-relevant metabolites descyanothiacloprid and desnitroimidacloprid. The pesticide metabolites showed a high potency, compared with their respective parent compounds. After this basic system characterization, the hitherto data-poor pesticides cycloxaprid and flupyradifurone were comparatively profiled in astrocytic and neuronal test systems. They showed the typical features of alpha7 nAChR agonists. The disruption of cholinergic signalling in astrocytes suggests that neonicotinoids affect not only neurons in human brains. Therefore, future neurotoxicity screening approaches may need to consider astrocyte toxicity. Full article

Background/Objectives: Germ cell tumors (GCTs) are thought to arise from primordial germ cells that fail to appropriately differentiate and instead retain pluripotency programs. PRDM14 is a key regulator of pluripotency and primordial germ cell specification and is aberrantly expressed in multiple GCT subtypes. However, the role of PRDM14 in GCT malignancy remains unclear. In this study, we investigated whether PRDM14 functions in GCTs through CBFA2T2, a transcriptional corepressor previously identified as a PRDM14-interacting partner in pluripotent stem cells and developmental models. Methods: To determine the presence and level of PRDM14 and CBFA2T2 in GCT, a panel of GCT lines were assessed for RNA and protein expression and interaction. Then, to better understand the biological effects of PRDM14 and CBFA2T2 within GCTs, PRDM14 and CBFA2T2 knockdowns were employed. Results: We show that PRDM14 and CBFA2T2 are expressed across GCT cell lines, colocalize predominantly in the nucleus, and cooperate as a complex in GCT cell lines. Knockdown of either PRDM14 or CBFA2T2 resulted in reduced expression of key pluripotency genes and a significant impairment of cell proliferation, indicating a shared role in maintaining an undifferentiated, proliferative state. Transcriptomic analysis following PRDM14 or CBFA2T2 depletion revealed extensive overlap in differentially expressed genes and convergent alterations in developmental and metabolic signaling pathways. Conclusions: Together, these findings suggest that PRDM14 and CBFA2T2 form a functional complex that sustains pluripotency and proliferation in GCT cells. This supports a model in which persistence of germline regulatory mechanisms contributes to GCT malignancy, highlighting this interaction as a novel component of GCT biology. Full article

Sepsis is a life-threatening condition characterized by a dysregulated host response to infection, leading to multi-organ dysfunction. Toll-like receptor signaling via MYD88- and TRIF-dependent pathways plays a central role in this process; however, its temporal and tissue-specific dynamics remain incompletely understood. The aim of this study was to investigate time-dependent transcriptional changes in MYD88- and TRIF-dependent signaling pathways across multiple organs in a murine model of sepsis. mRNA expression of MYD88, IRAK1, IRAK4, NF-kB, CCL4, CCL20, CCR2, IFN-β, IFN-γ, TNF-α, IL-1β, IL-2, IL-4, IL-8, IL-10, IL-18, Klotho, KLF4, HOXA5, NANOG and HIF1α was quantified using qRT-PCR in intestinal, kidney, liver and lung tissues at 24, 48, and 72 h following cecal ligation and puncture-induced sepsis in male C57BL/6J mice. Significant upregulation of innate immune signaling molecules, cytokines, chemokines, and interferon-related genes was observed in all tissues compared with controls. Genes associated with hypoxia and cellular regulation were also increased. These responses were tissue-specific and progressively intensified over time. Sepsis represents a dynamic, time-dependent, and tissue-specific process characterized by sustained activation of immune and hypoxic pathways, providing potential targets for time-stratified therapeutic strategies. Full article

Cardiovascular disease (CVD) is the leading cause of mortality worldwide. The healthy adult heart depends on flexible energy use, but a diseased or injured heart is associated with a loss of flexibility and metabolic remodeling. Since metabolism plays a central role in cardiac health and disease, there is a growing need to understand how metabolic reprogramming contributes to cardiac dysfunction and impaired CM maturation. Human-induced pluripotent stem-cell-derived cardiomyocytes (hiPSC-CMs) are widely used as a platform to study human cardiac development and disease mechanisms. However, current models are limited by metabolic and structural immaturity. This review provides an overview of the dynamic shifts in cardiac metabolic states from fetal development to senescence, while delineating the metabolic signatures of healthy versus disease states. These metabolic switches are orchestrated by a complex interplay of upstream signals driven by variations in substrate availability, post-translational modifications and key transcriptional regulatory networks, which ultimately regulate downstream cardiac remodeling and pathological cascades. As cardiac metabolic function is affected by a coordinated multicellular network, this review also includes the metabolic crosstalk between CMs and non-CMs, including fibroblasts, endothelial cells and immune cells. In addition, various strategies to further mature hiPSC-CMs are summarized to enhance their metabolic profiles. Investigating cardiac metabolic shifts bridges developmental biology, stem cell biology, and regenerative cardiology by revealing how energy metabolism governs cellular identity, maturation, and regenerative potential. These insights are essential for improving stem-cell-derived CMs for disease modeling, drug discovery, and heart repair. Full article