Search Results (387)

iPSC technology is well established in mammals but remains underdeveloped in non-mammalian species. A major barrier to generating avian iPSCs has been the lack of species-specific reprogramming factors and culture conditions capable of supporting self-renewal in avian pluripotent stem cells. Here, we report the generation of chicken iPSCs (ciPSCs) using a cocktail of seven chicken transcription factors (T7: Oct4, Sox2, Sox3, Klf4, c-Myc, Nanog, and Lin28B) combined with an optimized avian culture system. Transcriptomic and functional analyses identified Sox3, rather than Sox2, as the predominant SoxB1 factor in avian reprogramming. The resulting ciPSCs exhibited stable self-renewal for over 40 passages, expressed core pluripotency markers, differentiated into all three germ layers, and were transcriptionally similar to chicken ESCs. In chimera assays, ciPSCs contributed to somatic, extra-embryonic, and germline lineages, giving rise to gonadal PGC-like cells that did not acquire full germline competence. We further demonstrate that the T7 system generates iPSCs from quail, duck, peacock, zebra finch, and pigeon, and that duck iPSCs can form interspecies chimeras with donor cells detected in the host gonads. These findings establish a generalizable platform for avian iPSC generation with applications in developmental biology and germline preservation of endangered species. Full article

The landscape of human cardiac biology was transformed by the discovery that adult somatic cells can be reprogrammed into induced pluripotent stem cells, enabling patient-specific disease modeling, drug testing, and regenerative strategies without the prior ethical or biological constraints. Subsequent advances in directed differentiation made the generation of human iPSC-derived cardiac myocytes reliable and scalable. Despite this progress, a central limitation has remained: these cells are developmentally immature, resembling fetal cardiac myocytes in structure, metabolism, and function. This immaturity restricts their utility for modeling adult-onset disease, predicting drug responses, and achieving clinical translation. Maturation is now understood as a multifactorial symphony, requiring coordinated molecular, structural, and environmental inputs rather than single interventions. As a result, the field is shifting toward integrative approaches that incorporate 3D architecture, multicellular systems, and biomimetic environments to better replicate native cardiac tissue. While fully adult-like myocardium remains an ongoing goal, advances in bioengineering and system-level design are narrowing the gap, with success increasingly defined by the generation of functional cardiac tissue rather than isolated cell maturity. Full article

Human pluripotent stem cells (hPSCs) are a promising source for regenerative medicine due to their self-renewal and differentiation capacities. However, genetic instability acquired during reprogramming and in vitro culture presents major safety challenges for clinical translation. Recurrent mutations, especially structural variants (SVs), are of particular concern as they can impair differentiation and increase tumorigenic risk. In this review, we establish and systematically explore a central causal axis: SVs–three dimensional (3D) genome disruption–safety of hPSC-based therapy. We propose that SVs critically compromise therapeutic safety by perturbing the 3D architecture of the genome, leading to pathogenic rewiring of enhancer–promoter interactions. This rewiring, exemplified by “enhancer hijacking” and “enhancer loss,” can aberrantly activate oncogenes or silence tumor suppressors even in the absence of copy number variations. Thus, 3D genome disruption provides a key mechanistic explanation for SV-driven tumorigenic potential and impaired differentiation fidelity in hPSCs. By highlighting this causal axis, our review not only advances the mechanistic understanding of SV-associated risks but also provides actionable insights for the development of more rigorous quality standards for hPSC-based cell therapy products. Full article

Background: Treatment-resistant obsessive–compulsive disorder (OCD) is a heterogeneous and clinically challenging condition. Growing evidence suggests alterations in glutamatergic signaling within cortico–striatal–thalamo–cortical circuits, including those involving medium spiny neurons (MSNs), as well as genetic factors affecting synaptic organization, although the biological mechanisms underlying differential treatment response remain incompletely understood. Methods: This multicenter study presents a translational research framework aimed at investigating potential molecular and cellular correlates of treatment response in a cohort of patients with OCD, stratified according to their response to pharmacological treatments and transcranial magnetic stimulation (TMS). Peripheral blood mononuclear cells from clinically defined subgroups are reprogrammed into human induced pluripotent stem cells and differentiated into MSN-enriched neuronal cultures, enabling in vitro investigation of morphological, biochemical, and transcriptomic features associated with different clinical profiles. Optogenetic and pharmacological stimulation paradigms are applied to probe selected aspects of neuronal activation in vitro, providing a controlled and simplified experimental framework to explore cellular responses under different treatment conditions. By integrating clinical phenotyping with patient-derived cellular models, this study establishes a translational platform for hypothesis generation in the investigation of treatment response in OCD. Results and Conclusions: Preliminary clinical observations from an initial cohort undergoing neuromodulation are also reported to document feasibility and early clinical implementation of the study, providing an initial characterization of the cohort. Full article

Hair follicles (HFs) are highly accessible mini-organs that house diverse somatic and stem cell populations with broad therapeutic potential. In this study, we investigate the untapped utility of plucked HFs as a non-invasive tissue source for regenerative medicine. We demonstrate the successful isolation and expansion of keratinocytes and mesenchymal stem cells (MSCs) from plucked follicles using an enzyme-free explant culture method. HF-derived keratinocytes retained their epithelial identity and were efficiently reprogrammed into induced pluripotent stem cells (iPSCs). These iPSCs were further directed toward definitive endoderm and pancreatic progenitor fates, confirming their robust autologous regenerative capacity. Flow cytometric analysis of HF-MSCs validated a characteristic mesenchymal profile, and these cells exhibited classical trilineage plasticity alongside the ability to differentiate into dopaminergic neural progenitors. Furthermore, proteomic and vesicular characterization of the autologous HF secretome (aHFS) revealed a rich enrichment of regenerative cytokines and exosomes. The aHFS demonstrated potent wound-healing bioactivity in vitro. Collectively, these findings establish the plucked hair follicle as a highly practical, scalable source for both cell-based and cell-free therapies, highlighting the clinical value of early-stage follicular biobanking for personalized medicine. Full article

Adult neural stem cells (NSCs) maintain lifelong neurogenesis, a fundamental process for neuroplasticity, memory and brain homeostasis. Despite decades of research, translating basic NSC biology into effective clinical therapies remains a central challenge. Here we present a narrative review that provides a comprehensive update on the current landscape of adult NSC research, associating molecular mechanisms with the emerging translational technologies. First, we analyze the biological features and neurogenic sequences within canonical niches such as the subventricular lateral zone and the subgranular zone, emphasizing phylogenetic and migratory differences between rodent models and humans. Second, we integrate these mechanisms with the influence of environmental and pathological modulators, describing how aging, metabolic changes, chronic stress and neuroinflammation disrupt NSC quiescence and lineage progression. Finally, we highlight recent technological advances driving the field toward clinical applications. By examining current NSC isolation strategies, induced pluripotent stem cell modeling, direct somatic reprogramming and the use of CRISPR-Cas9-based gene-editing therapies, this review delineates the pathways to overcome existing methodological limitations. Ultimately, we provide an integrated context that connects the modulation of the neurogenic niches with advanced in vitro technologies, offering new perspectives for regenerative medicine and the treatment of neurological disorders. Full article

For over a decade, non-integrating Sendai virus vectors have been the gold standard for induced pluripotent stem cell (iPSC) reprogramming. However, as the field shifts toward regenerative and precision medicine and large-scale biorepositories, Sendai virus workflow necessitates dedicated viral-clearance testing, specialized manufacturing controls, and heightened regulatory oversight, leading to increased cost. While mRNA-based reprogramming offers a non-viral alternative, traditional mRNA delivery methods like electroporation are often physiologically disruptive. This study evaluates an mRNA-reprogramming platform that delivers lipid nanoparticles (mRNA-LNPs) via receptor-mediated endocytosis. By utilizing both Sendai virus and mRNA-LNP approaches to reprogram PBMCs from the same donor, we established a genetically identical starting point. Results demonstrate that mRNA-LNP-reprogrammed iPSCs maintain genomic integrity, retain the donor KCNH2 c.2398+5G>T variant, and exhibit characteristic colony morphology, pluripotency markers, and trilineage differentiation capacity consistent with the Sendai-reprogrammed counterparts. The mRNA-LNP-reprogrammed iPSCs differentiate into iPSC-derived cardiomyocytes presenting sarcomeric structures and electrophysiological activity, recapitulating a disease-specific phenotype. Notably, the mRNA-LNP workflow reached these milestones in significantly fewer passages than the Sendai virus workflow, markedly shortening timelines and reducing costs. These findings highlight mRNA-LNP reprogramming as a potentially attractive and effective, virus-independent platform to support future regenerative and precision medicine initiatives and scalable biobanking. Full article

The conversion of primed pluripotent stem cells to a naive-like state has emerged as a critical strategy for enhancing developmental potential and broadening applications in regenerative medicine. Conditioned media (CM)-based approaches provide a supportive microenvironment enriched with secreted factors that may facilitate this state transition without extensive genetic or chemical manipulation. In this study, we investigated the potential of human Wharton’s Jelly-derived mesenchymal stem cell-conditioned media (hWJ-MSCs-CM) and mouse embryonic fibroblasts CM (MEFs-CM) to support the conversion of primed rhesus monkey embryonic stem cells (rhESCs) into a naive-like state. The rhESCs were cultured under feeder-free and feeder conditions using both hWJ-MSCs-CM and MEFs-CM, exhibiting distinct morphological changes during conversion. Immunofluorescence analysis demonstrated the expression of pluripotency and naive markers under both conditions. Gene expression analysis further confirmed the upregulation of naive-specific genes and downregulation of primed markers, with statistically significant differences between groups. Additionally, epigenetic reprogramming was assessed, revealing differential effects of the CM sources on the reversion to a naive state. These findings highlight the potential of hWJ-MSCs-CM as a supportive system for naive-like state induction in primate ESCs. Full article

Cardiovascular disease remains the leading global cause of mortality, largely due to the limited regenerative capacity of adult human myocardium. Induced pluripotent stem cell-derived cardiomyocytes (iPSC-CMs) offer a scalable platform for cardiac repair and disease modeling; however, their persistent metabolic immaturity—characterized by reliance on glycolysis, reduced oxidative phosphorylation (OXPHOS), and structurally underdeveloped mitochondria—limits functional integration and long-term therapeutic efficacy. Recent advances indicate that targeted metabolic reprogramming can enhance mitochondrial biogenesis, increase ATP production, and improve stress resilience in iPSC-CMs. This review examines the complementary integration of CRISPR-based metabolic engineering and extracellular vesicle (EV)-mediated metabolic modulation as a systems-level strategy for cardiac maturation. We discuss CRISPR activation, interference, and epigenome-editing approaches targeting regulators such as PGC-1α, TFAM, and PPARs to promote stable enhancement of mitochondrial networks and respiratory capacity. In parallel, engineered EVs delivering miRNAs, metabolic enzymes, and redox modulators provide non-genomic mechanisms to optimize bioenergetic function and mitigate oxidative stress. By synthesizing mechanistic insights, quantitative bioenergetic metrics, and translational considerations, we propose CRISPR-EV synergy as a precision framework for durable metabolic maturation of iPSC-CMs, with implications for regenerative therapy, pharmacologic screening, and myocardial repair. Full article

The maturation of induced pluripotent stem cell-derived cardiomyocytes (iPSC-CMs) remains a major translational bottleneck in regenerative cardiac medicine, as current differentiation platforms yield electrophysiologically and metabolically immature phenotypes. This review explores emerging strategies to engineer “smart” biomaterial interfaces that actively instruct iPSC-CM maturation through synergistic biophysical and metabolic reprogramming. By integrating nanotopographical patterning, mechanoelectric coupling, and tunable substrate stiffness with metabolic interventions such as mitochondrial substrate optimization and fatty acid oxidation induction, the literature reveals consistent links between cell–matrix crosstalk, sarcomeric organization, calcium handling, and oxidative metabolism. Recent advances in bioactive scaffolds and extracellular vesicle (EV)-functionalized hydrogels are highlighted as platforms capable of approximating key features of the myocardium’s native electromechanical and bioenergetic environment. Across two- and three-dimensional culture systems, this review identifies recurring maturation patterns, persistent gaps in metric standardization and long-term phenotype stability, and ongoing limitations related to scalability and translational implementation. Collectively, the findings synthesized here indicate that convergence between biomaterial engineering and metabolic programming represents a critical design principle for advancing iPSC-CMs toward functionally mature, clinically relevant phenotypes. This integrated approach enhances the fidelity of iPSC-CMs for disease modeling, drug screening, and regenerative cardiac therapies. Full article

In multicellular organisms, the development of diverse cell types relies on stem cell differentiation through a hierarchy of fate decisions. Pluripotent stem cells first give rise to multipotent progenitors, which then undergo successive fate decisions to generate specialized cells within their respective lineages. Waddington used the metaphor of a marble rolling down a hill through hierarchically branching valleys that represent the various states of cell differentiation, with the final valleys at the bottom symbolizing the specialized cells. Mathematically, specialized cells are seen as stable attractors in a complex dynamical system that displays multistability. However, this framework does not necessarily describe the hierarchical branching of stem cell differentiation. In a recent study, we addressed this issue by assuming that each gene regulatory network (GRN) consists of hierarchically coupled gene subnetworks (modules) that are self-regulated due to epigenetic factors. Each module was modeled using the normal form of relevant bifurcations. Overall, this approach captures both multistability and hierarchical branching in differentiation. Here, the normal forms of bifurcations are replaced by realistic biochemical switches. Theoretical analysis and numerical simulations demonstrated that hierarchically coupled biochemical switches can depict the three fundamental aspects of Waddington’s epigenetic landscape: (a) differentiation trajectories exhibit hierarchical branching, (b) attractors are robust to perturbations (homeorhesis), and (c) the proportions of specialized cells are preserved. It was further shown that appropriate external interventions can induce either probabilistic cellular reprogramming or highly predictable reprogramming outcomes. The incorporation of biochemical switches, rather than purely abstract normal forms, can contribute to more biologically grounded mathematical models of stem cell differentiation. This work also highlights the importance of normal forms for qualitatively understanding cell state dynamics and for building realistic modular GRNs. Full article

Tail docking, serving as an important management intervention in animal husbandry, plays a significant role in regulating tail fat deposition and improving production performance and health status in fat-tailed sheep. This study systematically revealed the reprogramming effects of tail docking on the epigenetic landscape and transcriptome of fat-tailed sheep by integrating whole-genome bisulfite sequencing (WGBS) and RNA m6A methylated immunoprecipitation sequencing (MeRIP-seq). At the DNA level, the tail-docked group exhibited a pronounced trend of hypomethylation across multiple functional genomic regions, including promoters, exons, and introns. Differentially methylated regions (DMRs) were significantly enriched in pathways related to tissue development and stress response, such as the Hippo signaling pathway and adherens junctions. Pyrosequencing validation of the promoter region of the key gene DGAT1 further confirmed the reliability of the WGBS data. At the RNA level, RNA m6A modifications showed an overall up-regulated pattern: the tail-docked group displayed higher numbers of m6A peaks, greater total peak length, and increased genomic coverage compared to the control group, along with better overall prediction of modification sites. Genes associated with differential m6A peaks were closely related to processes such as stem cell pluripotency and cytoskeleton regulation. qPCR validation of several methylation-related enzyme genes (e.g., METTL3, FTO, YTHDF1) yielded results consistent with the sequencing trends. Through integrated analysis of DNA methylation and RNA methylation, we identified 143 genes with concurrent changes in methylation and mRNA expression, among which 41 genes were regulated by both DNA and RNA methylation. These genes were primarily enriched in the adherens junction pathway. Notably, two core genes CITED4 and ZNF644 showed significant changes across all three levels: DNA methylation, RNA methylation, and mRNA expression. This study systematically elucidates the epigenetic mechanism by which tail docking stress induces coordinated DNA hypo-methylation and RNA m6A hyper-methylation to regulate transcriptomic reprogramming in response to environmental intervention. The findings provide novel insights into the molecular basis of trait formation in livestock. Full article

As a type of cell with self-renewal ability and multi-directional differentiation potential, stem cells are closely related to their functions, such as reprogramming transcription factors, histone modifications, and energy metabolism. m⁶A (N⁶-methyladenosine modification) is one of the most abundant modifications in RNA, and dynamic reversible m⁶A modification plays an important role in regulating stem cell function. This review moves beyond listing isolated functions and instead adopts an integrated perspective, viewing m⁶A as a temporal regulator of cellular state transitions. We discuss how m⁶A dynamically regulates stem cell pluripotency, coordinates epigenetic and metabolic reprogramming, and serves as a central hub integrating key signaling pathways (Wnt, PI3K-AKT, JAK-STAT, and Hippo). Finally, using somatic reprogramming as an example, we elucidate the stage-specific role of m⁶A in complex fate transitions. This comprehensive exposition not only clarifies the context-dependent logic of m⁶A regulation but also provides a precise framework for targeting the m⁶A axis in regenerative medicine and cancer therapy. Full article

Late-onset sporadic Alzheimer’s disease (LOAD) is the most common form of dementia. The disease is characterized by progressive loss of memory and behavioral changes followed by neurodegeneration of all cortical areas. While the contribution of genetic and environmental factors is important, advanced aging remains the most important disease risk factor. Because LOAD does not naturally occur in most animal species, except humans, studies have traditionally relied on the use of transgenic mouse models recapitulating early-onset familial Alzheimer’s disease (EOAD). Hence, the development of more representative LOAD models through reprograming of patient-derived cells into neuronal, glial, and immune cells became a necessity to better understand the disease’s origin and pathophysiology. Herein, and focusing on neurons, we review current work in the field and compare results obtained with two different reprograming methods to generate LOAD patient’s neuronal cells: the induced pluripotent stem cell and induced neuron technologies. We also evaluate if these models can faithfully mimic cellular and molecular pathologies observed in LOAD patients’ brains. Full article