Stem Cell Research & Therapy最新文献

Background: Biliary atresia (BA) is a nongenetic cholangiopathy characterized by biliary obliteration. However, the underlying pathological mechanism remains unclear. We aimed to explore the epigenetic BA pathology by using BA-specific deciduous dental pulp stem cells (BA-SHED), which develop in parallel with cholangiocyte progenitor cells in human embryos.

Methods: BA-SHED were isolated from human exfoliated deciduous teeth of patients with BA using the colony-forming unit fibroblast method. After sequential stimulation with cytokines and chemicals in cultured BA-SHED, the in vitro bile duct-forming capacity was analyzed using quantitative reverse transcription polymerase chain reaction (RT-qPCR) and immunofluorescence. Expression of hepatocyte nuclear factor 6 (HNF6) and transforming growth factor beta receptor 2 (TGFBR2) was analyzed using immunoblotting and RT-qPCR. The regulation of chromatin architecture at the HNF6 promoter was analyzed using nuclease-accessible chromatin-qPCR and chromatin immunoprecipitation-qPCR.

Results: BA-SHED showed an inheritable increase in HNF6 levels, resulting in TGFBR2 suppression and deficiency in bile duct formation. BA-SHED also accumulated Brahma and P65 complexes around the HNF6 promoter with chromatin architecture remodeling. Tumor necrosis factor-alpha and interferon-gamma co-stimulation mimicked the epigenetic signatures of BA-SHED.

Conclusion: The present epigenetic memory in BA-SHED implies that BA-SHED imprint bile duct deficiency through TGFBR2 dysregulated by the HNF6 promoter activation epigenetically. Thus, BA-SHED are a potential model for expanding our knowledge in BA research.

Background: Evidence shows neural involvement in bone remodeling; regulating maxillofacial nerve repair modulates jawbone. Neural stem cell (NSC) therapy is limited by sources/ethics, but neural crest-derived dental mesenchymal stem cells (MSCs) like stem cells from the apical papilla (SCAPs) have strong neuroregenerative potential for NSC transdifferentiation. Schisantherin A (Sch-A), neuroprotective, enhances NSC proliferation/differentiation. This study explores optimal Sch-A concentration/duration for SCAP neural differentiation and effects on rat mental nerve repair/mandibular development.

Methods: SCAPs' mesenchymal stem cell properties were verified via flow cytometry and trilineage differentiation. Effects of different Sch-A concentrations were evaluated using CCK-8, colony formation, scratch assay, qRT-PCR, immunofluorescence, and Western blot. Transcriptome sequencing identified underlying mechanisms and determined optimal. A mental nerve injury model was established in 4-week-old SD rats (five groups; n = 4 per group) to assess neurorepair, functional recovery, and mandibular development following transplantation of Sch-A-induced SCAPs.

Results: Treatment with 10^- 9 mol/L Sch-A for 1 week induced robust neural differentiation in SCAPs, with high expression of nestin and NSE. Mental nerve-injured SD rats exhibited reduced lip sensation, abnormal nerve morphology, and inhibited transverse development of the anterior mandibular. Transcriptome analysis revealed Sch-A primarily acts via neuroactive ligand-receptor interaction pathway. Transplantation of induced SCAPs promoted nerve repair and restored mandibular development.

Conclusion: Sch-A at 10^- 9 mol/L concentration promotes the transdifferentiation of SCAPs into neural stem cell-like cells primarily through the neuroactive ligand-receptor interaction pathway. These Sch-A induced SCAPs effectively repair mental nerve injury and facilitate normal mandibular development.

Background: Hematopoietic stem cell transplantation (HSCT) is a cornerstone in the treatment of hematological disorders. However, its application is frequently complicated by acute and chronic graft-versus-host disease (aGVHD/cGVHD), pathological conditions in which donor-derived immune cells attack host tissues. With suboptimal survival rates and limited therapeutic options, GVHD remains a major clinical challenge. Mesenchymal stem cells (MSCs) have emerged as a promising therapeutic modality due to their immunomodulatory capabilities, yet standardized protocols for their use in preventing or treating GVHD have not been established.

Methods: We performed a comprehensive literature search of PubMed, Web of Science, EMBASE, and the Cochrane Library up to 10 February 2025 to identify eligible randomized controlled trials (RCTs). Study selection was based on the PICOS framework, and the risk of bias was assessed using appropriate quality appraisal tools. Outcome data were systematically extracted and synthesized via meta-analysis.

Results: A total of 15 RCTs were included. The meta-analysis revealed that MSC administration significantly reduced the incidence of aGVHD (OR: 0.47; 95% CI 0.32-0.71; p = 0.00003) and cGVHD (OR: 0.50; 95% CI 0.34-0.74; p = 0.0005) compared with controls. MSC therapy was also associated with improved response rates in steroid-refractory aGVHD (SR-aGVHD) (OR: 1.50; 95% CI 1.04-2.17; p = 0.03).

Conclusion: MSCs demonstrate efficacy in preventing both aGVHD and cGVHD following HSCT, particularly in moderate to severe forms. A dose range of 1 × 10⁶ to < 4 × 10⁶ cells/kg was associated with optimal prophylactic outcomes. For SR-aGVHD, MSC infusion resulted in significantly higher remission rates compared to conventional treatments, especially in severe cases.

Chronic traumatic encephalopathy (CTE), a progressive neurodegenerative disorder, poses a significant threat to human health. The lack of validated animal models has impeded mechanistic studies and the development of treatments for CTE. Recent evidence suggests that bone marrow mesenchymal stem cell-derived exosomes (BMSC-exos) represent a promising strategy for treating central nervous system injuries; however, their efficacy and mechanisms of action in CTE remain unexplored. In this study, we developed and optimized a CTE mouse model that recapitulates the core clinical features observed in CTE patients, including the delayed symptom onset. Using this model, we investigated the therapeutic effects of BMSC-exos. Our results indicate that BMSC-exos ameliorated anxiety-like behaviors and cognitive deficits in CTE mice, restoring them to levels comparable to those in noninjured control mice. Mechanistically, analysis of the hippocampal subgranular zone (SGZ) revealed that BMSC-exos restored the chronic CTE-induced reduction in the number of doublecortin (DCX)-positive immature neurons without altering the population of Sox2-Nestin-double-positive neural stem cells, indicating a primary effect on promoting neuronal differentiation efficiency or immature neuron survival rather than stem cell proliferation. Furthermore, BMSC-exos preserved neuronal structural integrity during late-stage CTE, indicating a critical role in maintaining synaptic plasticity and dendritic complexity. Collectively, our study provides promising evidence for the therapeutic potential of BMSC-exos in CTE, offering new insights for future CTE therapeutics.

Retinoblastoma (RB), which is the most common pediatric intraocular malignancy driven by RB1 inactivation, presents with clinical challenges, such as treatment toxicity, relapse, and resistance. Traditional models inadequately replicate human RB genetics or tumor heterogeneity, warranting the development of advanced in vitro platforms. Retinal organoids generated from human pluripotent or patient-specific stem cells enable three-dimensional(3D) modeling of the tumor microenvironment, drug screening, and mechanistic studies. This review summarizes RB pathogenesis, including RB1 loss, MYCN amplification, epigenetic dysregulation (e.g., METTL3-mediated m6A), and dysregulated pathways (PI3K/AKT/mTOR, Hedgehog), and highlights CRISPR-engineered organoids for identifying cone precursors as tumor origins and validating therapies (CDK4/6 inhibitors and sunitinib). Despite these advances, organoid applications are limited by high costs, variable success rates, incomplete immune/vascular mimicry, and limited scalability. Current microfluidic systems partially address vascularization but lack functional perfusion. Future efforts should integrate multiomics, refine vascularization via 3D bioprinting, and develop immunocompetent models to address the disparity between preclinical research and clinical application. Organoid technology has the potential to advance personalized therapies and ultimately enhance the survival and quality of life of patients with RB worldwide.

Background: Periodontal tissue regeneration can be achieved by periodontal ligament stem cells (PDLSCs) through its regulating the immune system. However, the specific signal or molecular mechanism remains unreported. The interaction between MSCs and macrophages (Mφ) has been the focus of the research in recent years. The objective of this study is to examine the effect of direct co-culture of human periodontal ligament stem cells (hPDLSCs) and macrophages on the osteogenic differentiation of hPDLSCs and the polarization of macrophages, and to explore the potential involvement of the EphB4/ephrinB2 signaling pathway in the interaction of co-cultured hPDLSCs and macrophages.

Methods: hPDLSCs isolated from human periodontal ligament were co-cultured with non-activated M0 macrophages (M0-Mφ) induced from THP-1. Quantitative reverse transcription polymerase chain reaction (qRT-PCR), alkaline phosphatase (ALP) staining and assay, as well as Alizarin red staining (ARS) were carried out to evaluate hPDLSCs osteogenic differentiation. qRT-PCR and Enzyme-Linked Immunosorbent Assay (ELISA) were employed to detect the expression of macrophage polarization-related factors. Western Blot was utilized to detect the expression of EphB4, ephrinB2, ERK1/2 and STAT3.

Results: When M0-Mφ was directly co-cultured with hPDLSCs at a ratio of 5:1, the co-culture system significantly promoted the osteogenic differentiation of hPDLSCs, as demonstrated by enhanced ALP staining/activity, ARS mineralization and upregulated mRNA expression of osteogenic markers (RUNX2, ALP, OCN/BGLAP, and OPN/SPP1). Meanwhile, the co-culture system markedly increased anti-inflammatory factor expression (TGF-β1 and IL-10) and decreased the pro-inflammatory factors (TNF-α and IL-1β), indicating enhanced polarization of alternatively activated macrophages (M2-Mφ). The mRNA and protein expression of EphB4 and ephrinB2 showed a significant increase with the time extension of the two cells' co-culture. However, pharmacological interruption of EphB4/ephrinB2 signaling pathway was associated with a decrease of hPDLSC osteogenic differentiation, M2 macrophage polarization, and p-STAT3 expression in the co-culture system.

Conclusions: Our data suggest a potential mediatory role for the EphB4/ephrinB2 pathway in the osteogenic differentiation of hPDLSCs and the polarization of M2-Mφ within the co-culture system. Its regulatory effect on the osteogenic differentiation of hPDLSCs may involve the STAT3 signaling pathway.

Background: Mesenchymal stem cells (MSCs) possess immunomodulatory properties and are concurrently influenced by the local inflammatory microenvironment. Extracellular vesicles (EVs) derived from MSCs (MSCs-EVs) replicate the therapeutic effects of their parent cells while avoiding the limitations of cell therapy. Elucidating the impact of diverse inflammatory factors on the composition and functionality of MSCs-EVs is crucial for their optimal use, though this remains incompletely understood. The aim of this study was to explore the immunomodulatory effects and underlying mechanisms of EVs derived from adipose-derived stem cells (ADSCs) pretreated with TNF-α and TGF-β1 (α-EVs and β-EVs) on macrophages and tissue-engineered cartilage regeneration.

Methods: Isolation and identification of EVs from ADSCs cultured in 3D spheres. The effects on chondrocyte and macrophage proliferation, migration, and polarization were evaluated in vitro. In vivo, chondrocytes-laden porous GelMA hydrogel with EVs were implanted into mice to assess chondrocyte regeneration and macrophage infiltration. Small RNA sequencing revealed distinct EVs-miRNA expression profiles, and the target validation confirmed the molecular mechanism.

Results: In vitro, both α-EVs and β-EVs demonstrated the capacity to modulate macrophage phenotypes. α-EVs more effectively reduced M1 macrophage markers and enhanced M2 polarization. Besides, β-EVs exhibited a stronger inhibitory effect on macrophage proliferation and migration, while also promoting chondrocyte proliferation and extracellular matrix (ECM) formation. In vivo, β-EVs significantly improved ECM deposition and chondrocyte maintenance, while both EVs groups reduced M1 infiltration and increased M2 presence. Small RNA sequencing identified miR-378a-3p upregulation in both α-EVs and β-EVs, targeting Signal-regulatory protein alpha (SIRPα) to modulate the immune status of macrophages.

Conclusions: Both TNF-α and TGF-β1 enhanced the immunomodulatory effects of EVs, with TGF-β1 showing a stronger capacity to promote chondrocyte proliferation and ECM synthesis. The miR-378a-3p/SIRPα axis was identified as a key mechanism underlying the protective effects of both α-EVs and β-EVs. This study provides valuable insights into optimizing EVs-based regenerative strategies to regulate the local inflammatory microenvironment and promote the regeneration of engineered tissues.

Background: Desmin-related cardiomyopathies caused by mutations in the DES gene are characterized by cytoskeletal disorganization and impaired cardiomyocyte function. Viral infections, particularly with Coxsackievirus B3 (CVB3), have been implicated as environmental triggers for cardiac decompensation. However, the interaction between desmin mutations and viral infection has never been explored.

Methods: Human induced pluripotent stem cell-derived cardiomyocytes (hiPSC-CMs) from a healthy donor (control-CMs) and from patients carrying DES^S46Y, DES^D214-E245del, or DES^P419H mutations (DES^mut-CMs) were infected with the cardiovirulent enterovirus CVB3/28. Structural changes were assessed by immunofluorescence for sarcomeric proteins and desmin. Contractile function was evaluated through video-based motion tracking. Viral replication, protein expression and antiviral responses were measured via plaque assays, immunostaining, and qPCR. Coxsackievirus and Adenovirus Receptor (CAR) and cell-surface vimentin expression were quantified post-infection.

Results: DES^mut-CMs exhibited baseline sarcomeric disorganization and desmin aggregation, which were further aggravated by CVB3/28 infection in a mutation-specific manner. CVB3/28 significantly reduced spontaneous contractility in control-CMs, DES^S46Y, and DES^P419H-CMs, with minimal effect in DES^D214-E245del-CMs. Infected DES^mut-CMs showed enhanced viral replication, increased VP1 expression and elevated virion release. This was accompanied by a stunted IFN-β response, reduced APOBEC3A expression, and infection-induced upregulation of viral receptors CAR and cell-surface vimentin.

Conclusion: CVB3/28 infection compromises the structural integrity and contractile function of cardiomyocytes and exerts a more severe effect in cells harboring DES mutations. These findings underscore a pathogenic synergy between genetic cytoskeletal defects and viral infection, revealing a mechanistic basis for the heightened vulnerability of patients carrying mutation in DES gene to virus-induced cardiac decompensation. CVB3/28 infection disrupts cardiomyocyte structure and impairs contractility, with more severe effects in cells carrying DES mutations. By enhancing viral replication and weakening antiviral defenses, DES mutations act synergistically with CVB3/28 infection to increase the risk of cardiac dysfunction.

Background & aims: Macrophages related inflammation plays a pivotal role in the progression of acute-on-chronic liver failure (ACLF). Our previous study has found that mesenchymal stem cells (MSCs) alleviate inflammatory damage in ACLF mice by promoting polarization of M2 macrophages through upregulating the expression of Mer tyrosine kinase (MERTK). In this study, we investigate the specific mechanism by which MSCs regulate MERTK.

Methods: Bioinformatics was used to predicted the candidate transcription factors for Mertk gene and FOS like antigen 1(FOSL1) was chosen. After overexpressing or knocking down Fosl1, MERTK, iNOS and Arg-1 were analyzed in Raw264.7 and/or J774a.1 cells. Conditioned medium (CM) of MSCs was cocultured with macrophages and the expressions of TGF-β1 and FOSL1 were detected. Male Balb/c mice aged 5-6 weeks were used to establish ACLF mice model. And adeno-associated virus or MSCs-CM was injected through tail vein. Then mouse liver tissue was collected and analyzed.

Results: FOSL1 promotes M2 polarization of macrophages by upregulating the expression of MERTK in vivo and in vitro. The luciferase reporter assays indicate that FOSL1 acts as a transcription factor of Mertk gene. Furthermore, MSCs-CM promotes expression of FOSL1 and M2 polarization of macrophages through TGF-β1 receptor. After knocking down TGF-β1 in MSCs using shRNA, shRNA-CM could not upregulate the expression of FOSL1, and promote M2 polarization of macrophages as CM does.

Conclusions: Our findings show that TGF-β1 secreted by MSCs promotes M2 macrophages polarization via FOSL1/MERTK axis in ACLF mice, providing a novel therapeutic target for ACLF treatment.

Background: Over the past decade, forward programming of human pluripotent stem cells (hPSCs) using various transcription factor (TF) combinations has been widely applied in neuroscience research. Ectopic NGN2 expression in hPSCs has been widely used for rapidly generating in vitro models of induced neurons (iNs) that are predominantly composed of excitatory glutamatergic neurons. Achieving a more balanced synaptic communication between excitatory and inhibitory neurons is essential for physiologically relevant in vitro models. Additionally, incorporating hPSC-derived astrocytes into models, rather than commonly used primary astrocytes, would more closely mimic in vivo disease phenotypes, especially for those associated with astrocyte dysfunction.

Methods: Inducible hPSC lines were generated by targeting the AAVS1 safe harbor site with TF transgene cassettes using CRISPR/Cas9. Forward programming was achieved through forced expression of NGN2 for glutamatergic neurons (iGlutNs), ASCL1/DLX2 for GABAergic neurons (iGABANs) and SOX9/NFIB for astrocytes (iAstros). Cell identity was validated by immunocytochemistry and bulk RNA sequencing. Functional properties were characterized on multielectrode arrays (MEAs).

Results: Bulk RNA sequencing confirmed lineage-specific differentiation while revealing distinct transcriptomic profiles between iAstros and human primary astrocytes. Functional assays demonstrated robust inhibitory control of network dynamics in co-culture with iGABANs on MEA, with enhanced responses to GABA_A receptor-targeting drugs including picrotoxin, bicuculline and clonazepam. Neurons co-cultured with iAstros showed reduced spontaneous activity compared to those cultured with primary astrocytes.

Conclusion: We successfully generated hPSC-derived excitatory and inhibitory neurons to establish an appropriate E/I balance in vitro, supported by primary astrocytes. Although astrocyte identity was confirmed in our hPSC-derived astrocytes, further optimization is required to achieve full functional maturation. This approach to developing an isogenic co-culture system derived from a single hPSC line may more faithfully replicate native neural network dynamics, offering a physiologically relevant platform for studying neurological disorders and screening therapeutic compounds.