Generating mature human induced pluripotent stem cell-derived cardiomyocytes (hiPSC-CMs) remains a major obstacle to accurate disease modeling and cardiac repair. As the transcription factor Irx3 is a key determinant of ventricular conduction system fate in mice, we hypothesized that suppressing IRX3 expression accelerates human working cardiomyocyte differentiation. Here, we demonstrate that depleting IRX3 enhances hiPSC-CM differentiation. IRX3-knockout (KO) hiPSCs generated a greater number of cardiomyocytes with elevated expression of TNNI1 and CX43. Notably, IRX3-KO cardiomyocytes exhibited improved electrophysiological properties, more uniform mitochondrial distribution, better sarcomere organization, and enhanced intercellular connectivity. We observed that IRX3 expression peaks during the early stages of cardiomyocyte differentiation, whereas IRX3-KO cardiac progenitors have increased expression of GATA4, NKX2-5, and TBX5, as well as enhanced cell proliferation. These integrative analyses indicate that IRX3 influences cardiomyocyte differentiation by modulating the gene regulatory networks driven by GATA4, NKX2-5, and TBX5, providing functional evidence linking gene regulatory networks to the structural and electrophysiological development of cardiomyocytes. Collectively, these findings identify IRX3 as a key regulator of early cardiac commitment and highlight the potential of IRX3 suppression to enhance the molecular and functional phenotype of hiPSC-derived cardiomyocytes.

Glioblastoma, the most aggressive and lethal form of brain cancer, is defined by profound genomic instability, with Chromosomal Instability (CIN) playing a central role in driving tumor progression, therapy resistance, and poor prognosis. CIN is characterized by numerical and structural alterations, is driven by mechanisms such as mitotic errors, centrosome amplification, spindle assembly checkpoint dysfunction, and defective DNA repair pathways. These aberrations contribute to tumor heterogeneity, leading to the emergence of Glioblastoma Stem Cells (GSCs) with enhanced plasticity, therapy resistance, and metastatic capacity. Chromothripsis, frequently involves specific chromosomes and stems from micronuclei rapture, resulting in chromosomal rearrangement. The immune implications of CIN are also critical, with the Cyclic GMP-AMP Synthase-Stimulator of Interferon Genes (cGAS-STING) pathway toggling between anti-tumor immunity and immune evasion. Therapeutic strategies targeting CIN are explored, including inhibitors of centrosomal clustering, DNA damage response pathways, and spindle assembly components, as well as innovative approaches like Chimeric Antigen Receptor T (CAR-T) cell therapies and nanoparticle-based drug delivery systems. Advances in single-cell sequencing provide transformative insights into CIN-driven glioblastoma heterogeneity and therapeutic vulnerabilities. By integrating mechanistic understanding with translational strategies, this review underscores CIN as both a therapeutic challenge and an opportunity, charting a path toward improving glioblastoma treatment outcomes and patient survival.

Cell-loaded magnetic microrobots hold promise for the targeted delivery of stem cells in tissue regeneration. However, achieving high cell-loading capacity alongside effective co-delivery of functional bioactive molecules remains a major challenge. Herein, we present a biodegradable magnetic microrobot featuring a core-shell architecture, designed to co-deliver stem cells and bioactive molecules. These microrobots are fabricated using a rotation-induced inertial focusing technique, which allows control over their size and morphology while significantly enhancing cell-loading capacity. The core, composed of a biodegradable magnetic microsphere (MMS), enables external magnetic navigation and sustained drug release, while the outer shell, formed by mesenchymal stem cells (MSCs), ensures therapeutic viability. The microrobots demonstrate robust magnetic maneuverability and spatial control even in complex and viscous environments. We validated their efficacy in both a rabbit in vivo model and an ex vivo human cartilage model, where co-delivery of MSCs and transforming growth factor-β1 (TGF-β) led to significant improvements in cartilage repair and functional tissue regeneration. This biodegradable core-shell microrobot offers a scalable, multifunctional platform for efficient co-delivery of cells and bioactive molecules, with strong potential for clinical translation in regenerative medicine.

Complement factor H (CFH) is an immune regulator that inhibits the complement system. Here, we identify CFH as a secreted and regulated factor in human bone marrow skeletal stem cells (hBMSCs) culture during osteoblast (OB) differentiation. To explore its role in bone formation, we investigated the effects of CFH on OB differentiation and bone homeostasis. CFH-deficient hBMSCs exhibited impaired OB differentiation, whereas CFH overexpression or supplementation enhanced OB differentiation in hBMSCs. In vivo, CFH-deficient (CFH-/-) mice displayed reduced trabecular and cortical bone mass, decreased bone formation, and diminished bone strength. Primary CFH-/- mouse bone marrow MSCs (mBMSCs) showed reduced osteogenesis but enhanced osteoclast differentiation, consistent with lower serum levels of the bone formation marker P1NP and elevated bone resorption markers TRAPc and CTX-1. Furthermore, CFH-/- mice exhibited delayed fracture healing and accelerated bone loss following ovariectomy or high-fat diet feeding. Clinically, CFH levels in bone marrow plasma were negatively correlated with fracture risk in patients. Notably, plasma CFH levels were positively associated with bone mineral density (BMD) and were significantly reduced in patients with osteoporosis. These findings establish CFH as a key regulator of osteogenesis and bone homeostasis, with potential implications for bone-related disorders.

Charcot-Marie-Tooth disease type 2A (CMT2A) is the most common axonal CMT and is associated with an early onset and severe motor neuropathy. CMT2A is mainly caused by dominant mutations in the MFN2 gene, encoding mitofusin-2, a GTPase located in the outer membrane of the mitochondria and endoplasmic reticulum (ER). Mutations in MFN2 affect mitochondrial dynamics. We previously demonstrated that mutated MFN2 further disrupts contacts between the ER and the mitochondria, leading to axonal degeneration. There are no treatments for CMT2A, and those currently under development primarily focus on restoring mitochondrial function. Here, we provide proof of concept that neuronal overexpression of wild-type MFN2 (MFN2WT) provides therapeutic benefit in transgenic CMT2A mice as well as in CMT2A-motor neurons derived from induced pluripotent stem cells. Intrathecal delivery of an AAV9 vector expressing MFN2WT effectively targets motor and sensory neurons, restoring ER-mitochondria contacts and mitochondrial morphology, thereby preserving both neuromuscular junction integrity and motor function. Strikingly, therapeutic efficacy is also achieved by administering the vector after the onset of symptoms. Importantly, AAV administration was well tolerated, with no evidence of hepatotoxicity or dorsal root ganglion inflammation. We further show that CMT2A pathology can be corrected in vitro and in vivo using an ER-targeting MFN1 isoform that selectively enhances ER-mitochondria contacts. These results establish that restoring contacts between the ER and mitochondria using gene therapy is a promising therapeutic avenue for CMT2A.

Transcription factor (TF) upregulation accompanies many cellular state transitions, yet how increased TF abundance impacts gene regulation remains unclear. Two broad models are often invoked, whereby higher TF levels amplify the expression of preexisting target genes, or, by mass-action binding, expand genome engagement and regulation to lower-affinity sites. We sought to elucidate how these two regulatory modes contribute to cell differentiation in a well-characterized myogenic system by upregulating the expression of the myogenic TF MyoD1 in C2C12 myoblasts. Unexpectedly, elevated MyoD1 levels impaired myoblast fusion (a hallmark of myogenic differentiation), yet enabled robust contraction in myotubes that did form. Live-cell single-molecule imaging and CUT and RUN profiling revealed that elevated MyoD1 dosage increased total genome-wide chromatin binding and broadened genome occupancy by preferentially engaging lower-affinity sites. Integrating CUT and RUN with RNA sequencing (RNA-seq) experiments linked expanded MyoD1 binding to upregulation of cell adhesion genes. Cell mixing and fractionated RNA-seq experiments supported a two-population model in which an adhesion-gene-upregulated, unfused myoblast population supported contraction of myotubes formed by fusion-competent cells. Ectopic expression of several individual MyoD1-upregulated cell adhesion genes was sufficient to recapitulate the "off script" myotube contraction phenotype. Together, these results support a MyoD1 dose-dependent "spillover" model, in which increased TF abundance broadens cis-regulatory engagement and produces distinct cell differentiation outcomes.

Duchenne muscular dystrophy (DMD) is a debilitating and fatal X-linked disease affecting 1/5,000 males worldwide that currently has no cure [D. Duan, N. Goemans, S. Takeda, E. Mercuri, A. Aartsma-Rus, Nat. Rev. Dis. Primers7, 1-19 (2021), 10.1038/s41572-021-00248-3]. Vast amounts of research have been conducted on DMD, and one of the most common animal models for DMD studies is the mouse muscular dystrophy (mdx) model [J. W. McGreevy, C. H. Hakim, M. A. McIntosh, D. Duan, DMM Dis. Model. Mech.8, 195-213 (2015), 10.1242/DMM.018424/-/DC1]. Unfortunately, despite its shared genetic etiology, the mdx mouse shows a relatively mild dystrophic phenotype compared to affected humans, limiting its overall utility as a research model (G. Donen, N. Milad, P. Bernatchez, J. Neuromuscul. Dis.10, 1003 (2023), 10.3233/JND-230126]. Notably, mdx mice have a mutation preventing the production of full-length dystrophin but are still able to produce numerous short isoforms of dystrophin. Here, we provide a comprehensive functional characterization of DMD-Null mice, which lack all dystrophin isoforms. Our studies demonstrate that DMD-Null mice show a more severe skeletal muscle phenotype than mdx mice, characterized by profound weakness, decreased exercise tolerance, and impaired muscle regeneration, while utrophin upregulation was similarly observed in DMD-Null and mdx mice. We identify a marked deficit in satellite cell proliferation and myogenic differentiation, accompanied by downregulation of regenerative gene programs. These findings suggest potential contributions of short dystrophin isoforms to muscle stem cell function, and establish DMD-Null mice as a unique model for investigating the pathogenesis of DMD and testing therapeutic interventions targeting satellite cell health and regeneration.

A pseudostratified epithelium lines the upper airways and is maintained by stem cells that express a basal cell phenotype. These airway stem cells share molecular and functional similarities with other tissue stem cells, particularly those that are involved in development or postnatal maintenance of the esophagus and epidermis. Thus, analysis of gene function in the airway is complicated by the potential for lethal non-respiratory phenotypes. Conditional genetic approaches employing cell type-specific and/or temporally controlled expression systems provide some options, but limitations remain. To overcome these concerns, we tested a novel microsurgical approach in which a chimeric airway is created by orthotopic transplantation of transgenic mouse tissue into the trachea. We hypothesized that precise temporal and spatial control of gene expression in the pseudostratified tracheal epithelium would prevent life-threatening collateral tissue injury and allow analysis of essential genes. First, we demonstrated that the graft was revascularized allowing parenteral administration of tamoxifen. Second, we transplanted tracheal tissue from Krt5CreERT2; Rosa26mTmG or Col1a2CreERT2; Rosa26mTmG donors into wild type mice and established that tamoxifen-dependent recombination in basal cells or fibroblasts was highly efficient and restricted to the graft. Finally, we assessed the feasibility of knocking out an essential gene, Itgb1, by transplanting tracheal segments from Krt5CreERT2; Rosa26mTmG; Itgb1flox/flox transgenic mice into wild type mice. We demonstrated highly selective recombination in the graft and long-term survival. We conclude that the chimeric airway model allows analysis of essential gene function in the airway and has the potential to be a versatile tool for preclinical testing of targeted gene therapies.

Spermatogonial stem cells (SSC) are of high significance in animal reproduction, breeding, and regenerative medicine. Cryopreservation of putative SSC is a prerequisite for long-term storage and future applications. However, dissociated putative SSC are very susceptible to cryostress and undergo apoptosis, thereby reducing their functional competence. To minimize the dissociation-induced apoptosis during cryopreservation and post-thaw culture, the role of anti-apoptotic molecule, Y-27632, was evaluated in sheep putative SSC.

To evaluate whether intraosseous (IO) autologous bone marrow concentrate (BMC), containing mesenchymal stem cells (MSCs) enumerated in vitro as colony forming unit-fibroblasts (CFU‑F), reduces or delays conversion to total hip arthroplasty (THA) compared with matched conservative care, and to assess the influence of disease progression and CFU‑F dose on outcomes.

Wound healing still faces many significant challenges, such as excessive oxidative stress, persistent inflammation and disrupted angiogenesis. To overcome these obstacles, we developed a biomimetic hybrid regulatory platform (ApoEVs@FeSe2-RGD) by anchoring redox-active FeSe2 nanorods onto mesenchymal stem cell-derived apoptotic extracellular vesicles (ApoEVs), followed by surface functionalization with cyclic RGD (cRGD) peptides. This regulator effectively synergizes the robust reactive oxygen species (ROS)-scavenging capacity of FeSe2 with the exceptional biocompatibility of cRGD-functionalized ApoEVs. In vitro assessments revealed that ApoEVs@FeSe2-RGD efficiently cleared abnormally produced ROS to recalibrate the oxidative microenvironment. Consequently, it inhibited pro-inflammatory activation while driving macrophage polarization toward an anti-inflammatory phenotype. The reparative cytokines derived from these reprogrammed macrophages significantly enhanced the migration, proliferation, and angiogenic capacity of endothelial cells. Furthermore, the in vivo assessments in a full-thickness wound healing model demonstrated that this vesicle-scaffolded regulator enhanced the wound closure rate and promoted re-epithelialization, collagen deposition, and neovascularization with good biosafety. Mechanistically, transcriptomic profiling combined with protein validation demonstrated that these therapeutic effects were mediated by the suppression of the NF-κB signaling pathway. Collectively, our study demonstrated that ApoEVs@FeSe2-RGD holds potential as a therapeutic strategy for wound healing by restoring redox homeostasis and orchestrating immune-vascularization coupling for high-quality tissue regeneration.

It has been 20 years since the pioneering work of Shinya Yamanaka and Kazutoshi Takahashi at Kyoto University led to the first successful generation of induced pluripotent stem cells (iPSCs) from mouse embryonic and adult fibroblast cells. iPSCs have the capacity to differentiate into any type of cell in the human body, and as such, they have become ubiquitous in medical and biological research. Throughout the development and use of iPSCs and their derivatives, fluorescent microscopy has been particularly integral, playing a central role in the characterisation of cells and analysis of cellular function of both iPSCs and their derived cells. On the 20th anniversary of their discovery, this short review summarises the development of iPSCs, their application in research and clinical settings, and highlights advances in microscopy and imaging methodologies that have been crucial in developing and characterising iPSCs and their derived cell types.

The rational design of hybrid nanocomposites that bridge nanoscale functionality and macroscale processability is central to advancing multifunctional materials for biofabrication and regenerative medicine. Craniofacial bone regeneration demands materials that provide mechanical stability while instructing stem-cell behavior toward functional tissue formation. Here, we present a rationally designed nanocomposite hydrogel integrating gold nanoparticles (AuNPs) functionalized with polyethylene glycol (PEG) and glutathione (GSH) into a gelatin methacrylate (GelMA) matrix to couple rheological tunability with biological activity. AuNPs were synthesized via chemical reduction, PEGylated for colloidal and thermal stability, and conjugated with GSH to introduce bioactive thiol and carboxyl groups that promote osteogenic signaling. Comprehensive physicochemical analyses confirmed successful functionalization, uniform dispersion, and stability across physiological conditions. Incorporation of PEG-GSH-AuNPs at optimized concentrations enhanced the complex viscosity, yield stress, and print fidelity of GelMA while preserving its shear-thinning behavior essential for extrusion-based bioprinting. Periodontal ligament stem cells (PDLSCs) encapsulated within the printed constructs maintained >90% viability and exhibited pronounced mineralization and upregulation of osteogenic markers, confirming the instructive potential of the hybrid bioink. This study establishes a modular strategy that bridges nanoscale material design with macroscale biofabrication, yielding a thermally stable, biologically active hydrogel capable of directing stem-cell fate. The resulting nanocomposite platform offers broad potential for precision bioprinting and next-generation bone and craniofacial tissue regeneration.

Advancements in 3D bioprinting demand bioinks that demonstrate not only precise printability and mechanical robustness but also consistent biochemical and cellular performance. From previous iterations in our laboratory, a silk fibroin-gelatin (SF-G) bioink was conjugated with mushroom tyrosinase (MT) as the enzymatic crosslinker. However, its clinical translation was hindered by several limitations, including batch-to-batch variation in concentration and enzymatic activity, protracted gelation kinetics, and inconsistent rheological and structural characteristics. To overcome these challenges, we developed a next-generation SF-G bioink employing horseradish peroxidase (HRP) and hydrogen peroxide (H2O2) as an alternative enzymatic crosslinking system. This approach facilitated rapid and tunable crosslinking through β-sheet enhancement, yielding hydrogels with improved stiffness, shape fidelity, and resistance to enzymatic degradation. Detailed rheological analysis confirmed optimal shear-thinning behaviour and print fidelity, while reactive oxygen species (ROS) quantification ensured cytocompatibility across physiologically relevant concentrations. Human bone marrow-derived mesenchymal stem cells (hBMSCs) encapsulated within the constructs maintained high viability and demonstrated robust osteogenic and chondrogenic lineage commitment. Furthermore, supplementation with triiodothyronine (T3) and transforming growth factor-β3 (TGF-β3) augmented matrix deposition and tissue-specific morphogenesis. Notably, a 10 U HRP-H2O2 formulation emerged as the most promising candidate, offering a strategic balance of mechanical integrity, bioactivity, and reproducibility. This optimized enzymatic system paves the way for scalable and clinically viable SF-G bioinks tailored for advanced tissue engineering and regenerative medicine applications.

After injury, the repair of brain tissue is hindered by poor neuronal regeneration and severe neuroinflammation. Although magnetoelectric (ME) stimulation has emerged as a promising wireless strategy to promote axonal growth, its efficacy is strictly dependent on the ME coupling efficiency of the materials used. Furthermore, while microglia-mediated inflammation remains a critical barrier to neurogenesis, integrating neuroprotective strategies to modulate this inhibitory environment with ME stimulation strategies has seldom been explored. Herein, we developed a multi-functional biomimetic hydrogel platform combining neuroprotective and neuroinductive properties simultaneously. This platform is composed of a collagen/oxidized hyaluronic acid-metformin hydrogel matrix with encapsulated biocompatible core-shell ME nanoparticles (NPs), MnFe2O4@Ba0.85Ca0.15Ti0.9Zr0.1O3 (MFO@BCZT), exhibiting a high ME coefficient. We confirmed that the metformin in the hydrogel effectively inhibited lipopolysaccharide (LPS)-induced M1 polarization of BV2 microglia, providing neuroprotection. Concurrently, under pulsed magnetic field stimulation, the embedded MFO@BCZT NPs generated local electrical signals, significantly promoting neuronal differentiation of neural stem cells (NSCs). In a rat traumatic brain injury (TBI) model, this synergistic system alleviated neuroinflammation, recruited endogenous NSCs, and promoted their neuronal differentiation and maturation within the lesion. This led to enhanced axonal regeneration, remyelination, synaptic reconstruction, and ultimately, improved cognitive function recovery. Overall, this work presents a promising strategy that combines ME stimulation-driven neuroinduction with metformin-mediated neuroprotection for effective brain tissue repair, shedding light on innovative strategies for biomaterial design and the treatment of central nervous system injuries.

Folic acid, a water-soluble B vitamin, is well known for its critical roles in neural tube development and its contributions to neonatal gut maturation and overall gut health. However, the specific mechanisms by which folic acid influences the intestinal mucosa remain incompletely understood. In this study, we aimed to explore the effects of folic acid on the proliferation and differentiation of intestinal stem cells (ISCs). A mouse model of intestinal mucosal injury was established by intraperitoneal injection of 5-fluorouracil (5-FU) at 50 mg per kg body weight once daily for five consecutive days. Our results demonstrated that folic acid enhanced epithelial barrier integrity and modulated epithelial function through the upregulation expression of tight junction components and nutrient transporters. Additionally, folic acid promoted the differentiation of intestinal epithelial cells and accelerated ISC renewal. Mechanistically, folic acid increased the expression of β-adrenergic receptors, whereas its stimulatory effects on enteroid growth and budding were attenuated by the β-adrenergic receptor antagonist propranolol, suggesting the involvement of β-adrenergic receptor signaling in folic acid-mediated epithelial regeneration. Collectively, these findings indicate that folic acid enhances ISC-driven epithelial regeneration in a β-adrenergic signaling-dependent manner, providing new insights into intestinal homeostasis and potential therapeutic strategies for mucosal injury.

Breast cancer (BC) remains the most prevalent and deadly malignancy among women worldwide. While photothermal therapy (PTT) and chemodynamic therapy (CDT) offer tumor-specific and minimally invasive advantages, their standalone efficacy is limited. Ligustilide (LIG) can exert antitumor effects by generating •O2-, but its bioavailability is poor, thus necessitating improved delivery strategies. Here, we reported a multifunctional nanoplatform (FeD@LIG) integrating PTT, CDT, and chemotherapy for the simultaneous elimination of breast cancer stem cells (BCSCs) and non-BCSCs. FeD@LIG was synthesized via one-step oxidative polymerization of dopamine, co-loading Fe3+ and LIG. The nanospheres exhibited excellent photothermal conversion efficiency (52.8%) and pH/GSH-responsive drug release. Upon near-infrared (NIR) irradiation, FeD@LIG triggered ROS generation via Fenton-like reactions and elevated local temperature, leading to enhanced oxidative stress, apoptosis, and LIG release. In vitro, FeD@LIG reduced cell viabilities of MCF-7 and MDA-MB-231 cells to 12.5 and 7.9%, respectively, and significantly inhibited BCSCs' self-renewal. In vivo, FeD@LIG achieved a tumor inhibition rate of 92.8% with minimal toxicity. Mechanistic studies revealed activation of the BAX/BCL-2/Caspase-3 pathway and suppression of Hedgehog signaling. Overall, FeD@LIG represents a promising nanotherapeutic strategy for overcoming drug resistance and recurrence by inhibiting both BCSCs and non-BCSCs through multimodal synergistic therapy.

Human Idiopathic Pulmonary Fibrosis (IPF) is a progressive and fatal lung disease with unknown etiology and lacking efficient treatments. Here, we reported that human urine stem cells (hUSCs) significantly alleviated pulmonary fibrosis via inhibiting macrophage-myofibroblast transition (MMT), which was identified as a pivotal pathological process in IPF, with the strong interaction among infiltrated macrophages, damaged alveolar epithelial cells, and myofibroblasts via single-nucleus RNA sequencing data analysis and co-immunostaining. In addition, hUSCs significantly alleviated pulmonary fibrosis by attenuating alveolar epithelial cell damage, reducing monocyte-derived macrophage infiltration, and suppressing MMT in the bleomycin-induced pulmonary fibrosis mouse model. Furthermore, we demonstrated hUSCs inhibited monocyte recruitment and MMT via paracrine actions in the macrophage-alveolar epithelial cell co-culture system. Mechanistically, DKK1, which was highly secreted by hUSCs and identified by Venn diagram analysis between the luminex assay in supernatants of THP1 treated with hUSC-CM and antibody array of hUSC-CM, might contribute to preventing MMT via suppressing the Wnt/β-catenin signaling pathway in macrophages. In summary, hUSCs exerted multifaceted protective effects against pulmonary fibrosis, at least in part through paracrine mechanisms involving DKK1 and its modulation of Wnt/β-catenin-associated fibrotic responses in MMT. Therefore, hUSCs might provide a potential therapeutic strategy for IPF clinically.

Neurogenesis in adult mammalian brain persists in restricted areas, especially the subgranular zone (SGZ) of the hippocampus and the ventricular-subventricular zone (V-SVz), where neural stem cells (NSCs) occupy neurogenic niches. These NSC niches provide signals that regulate stem cell behavior. Among extrinsic modulators, Growth Differentiation Factor 11 (GDF11 or BMP11) which is a transforming growth factor-β (TGF-β) superfamily member, was shown to play key role in the NSC biology and brain aging. In this review, the most recent molecular mechanisms of GDF11 signaling in the regulation of NSC will be addressed. GDF11 plays mainly through activin type II receptors (ActRIIA/B) and ALK4/ALK5, activating classical Smad2/3 pathways that impact transcriptional networks controlling neural cell behavior. Moreover, GDF11 stimulates non-Smad signaling pathways - including ERK, p38, JNK, and PI3K/AKT - providing context-dependent integration of proliferative and anti-proliferative signals. Furthermore, GDF11 functions as a feedback regulator limiting the number of progenitor cells and organizing neurogenic timing. In the adult brain, GDF11 plays important role in neurovascular remodeling, glial inflammatory states, and extracellular matrix interactions. Despite its recognized roles, the effect of GDF11 on aging remains a subject of intense debate, characterized by conflicting reports regarding its circulating levels, tissue-specific dynamics, and dose-dependent effects. Recent evidence suggests that GDF11 acts as a context-dependent modulator, integrating systemic, vascular, and cellular cues to maintain NSC homeostasis and neurogenic potential. Therefore, elucidating the exact cellular and molecular mechanisms by which GDF11 controls NSC behavior is vital to advancing novel therapeutic strategies for neurodegenerative disorders and age-related cognitive decline.

Human mesenchymal stem cells (hMSCs) undergo progressive functional decline during long-term ex vivo expansion, which limits their therapeutic potential. However, the contribution of intercellular adhesion to this process remains unclear. By comparing hMSCs at different passage stages, we found that replicative senescence is accompanied by impaired collective motility homeostasis in near-confluent monolayers, diminished traction forces and altered monolayer stress distribution, concomitant with upregulated N-cadherin expression. Notably, N-cadherin knockdown or pharmacological blockade of its homophilic binding using ADH-1 restored migratory dynamics, enhanced traction generation and alleviated senescence-associated phenotypes. These findings identify N-cadherin as a crucial regulator of replicative senescence and highlight intercellular adhesion as a potential target for delaying senescence during ex vivo stem cell expansion.