Stem Cell Research & Therapy最新文献

Background: Niosomes represent a promising non-viral gene delivery system, offering an alternative to viral vectors for the genetic modification of hard-to-transfect cells, such as mesenchymal stem cells (MSCs), which are pivotal in regenerative medicine. Specifically, SOX9 gene transfer is a valuable strategy for cartilage tissue repair, as it promotes chondrocyte differentiation while repressing hypertrophic and osteogenic markers. In this study, we investigated the potential of niosomes to deliver SOX9, using both parental and minicircle plasmids, to induce chondrogenic differentiation in primary bone marrow-derived human MSCs (hMSCs).

Methods: Niosomes were synthesised using the thin-film hydration method and complexed with either parental or minicircle SOX9 plasmids to form nioplexes. Physicochemical properties of niosomes and nioplexes were studied in terms of size, zeta potential, complexation, and protection capacity. Primary hMSCs were transfected in a 2D monolayer and 3D aggregate cultures using Lipofectamine as a positive control of transfection. Chondrogenic differentiation was assessed by gene expression (SOX9, ACAN, COLII, COLI, COLX), histological and immunohistochemical staining (Toluidine blue, haematoxylin & eosin and SOX9, COLII, COLI, COLX, respectively), and biochemical (proteoglycans, DNA and protein contents) analyses of main cartilage markers.

Results: SOX9 delivery via DP20CQ niosome systems significantly enhanced the expression of key chondrogenic markers (SOX9, ACAN, and COLII) and increased production of a characteristic hyaline-like cartilage matrix. In contrast, Lipofectamine-based complexes induced hypertrophic and fibrocartilaginous phenotypes, evidenced by increased expression of COLX and COLI. Quantification of proteoglycan production, along with proteins and DNA content, supported these findings. Both plasmid types promoted comparable chondrogenic outcomes, but parental plasmids yielded more consistent results than minicircles.

Conclusions: Delivery of SOX9 plasmids via niosomes promotes enhanced chondrogenic differentiation of primary hMSCs in a 3D aggregate culture system, leading to the formation of hyaline-like cartilage tissue. This non-viral strategy represents a promising gene delivery platform for cartilage reparative therapies.

Mesenchymal stem cells (MSCs) are multipotent stem cells with critical functions, including immunomodulation, multidirectional differentiation, anti-inflammatory activity, tissue repair, and regeneration. Recent studies demonstrate that MSCs can enhance hematopoietic stem cell engraftment, mitigate graft-versus-host disease (GVHD), address transplant-related complications, and treat conditions such as immune thrombocytopenia (ITP) and severe aplastic anemia (SAA). These therapeutic effects are largely attributable to the immunomodulatory and anti-inflammatory properties of MSCs. However, in hematologic malignancies, MSCs can exert both pro-tumor and anti-tumor influences. Exosomes, which are extracellular vesicles derived from MSCs (MSC-EVs), not only replicate many MSC functions but also exhibit greater chemical stability and lower immunogenicity. These characteristics make MSC-EVs particularly significant in the context of hematopoietic stem cell transplantation (HSCT). This review provides a detailed overview of the roles and clinical applications of MSCs in hematologic diseases, the properties of MSC-EVs, and their emerging significance in HSCT.

Background: Mesenchymal stem cells (MSCs) undergo senescence after expansion and in vitro culture under oxidative stress, which limits their clinical application. Ginsenoside Rh2 has been confirmed to regulate mitochondrial function, but its role in modulating the senescence of MSCs has not been clearly investigated.

Purpose: This study aims to explore the effects and underlying mechanisms of Rh2 in inhibiting the senescence of MSCs.

Methods: Transmission electron microscopey (TEM) and fluorescence staining assays were used to monitor changes in mitochondrial and lysosomal morphology and function in Rh2-treated senescent MSCs. Quantitative reverse transcription polymerase chain reaction (qRT-PCR) and Western blot analyses were performed to evaluate the expression levels of senescence-related cytokine genes and proteins.

Results: Rh2 can inhibited the senescence of MSCs by activating Sirtuin 1(SIRT1). At the molecular level, SIRT1 regulated the Pink1/Parkin-mediated mitophagy pathway and suppressed the secretion of senescence-associated cytokines (IL-6 and IL-8). Additionally, Rh2 influenced lysosomal stability and sultimately inhibited exosome secretion through direct activation of SIRT1.

Conclusion: These findings provide a potential strategy for using Rh2 to overcome the senescence of MSCs, thereby enhancing their clinical application.

Background: Vision loss due to retinal and corneal cell degeneration is significant clinical challenge, with current cell therapies hindered by limited donor cells. To address this, we developed a streamlined platform using human induced pluripotent stem cells (hiPSCs) that integrates 2D and 3D culture techniques to simultaneously generate retinal ganglion cells (RGC) and corneal lineages.

Methods: The non-integrated hiPSCs were used to ocular cells differentiation, four time-points cells were collected for 10×Genomics sing-cell RNA sequencing to trace RGC and corneal lineage development. The confirmed differentiated window phase to pick optic vesicles for 3D ocular organoids culture, and as well as directional induced corneal epithelium in remaining 2D dish. After FACS-based cell sorting, the enriched RGC-like and corneal cells were propagated in vitro, and these cells were transplanted into optic nerve crush (ONC) and corneal damaged mice respectively to observe the regenerative repairment capacity.

Results: Through single-cell RNA sequencing, we mapped differentiation trajectories and identified surface markers-CD184 and CD171 for RGCs, and CD104 for corneal progenitors facilitating purification. In mouse models, transplanted hiPSC-derived CD184⁺CD171⁺ RGC-like cells integrated into injured retinas, enhanced host RGC survival, and restored visual function following optic nerve injury. Concurrently, hiPSC-derived CD104⁺ corneal progenitor cells exhibited self-renewal, differentiation capabilities, and accelerated corneal repair with reduced neovascularization. Additionally, this platform enables the synchronous production of retinal and corneal organoids, which are valuable for both regenerative therapy and disease modeling.

Conclusions: Our study establishes a cost-effective surface marker-based method for deriving transplantable RGC and corneal lineage cells from hiPSCs, overcoming key obstacles in ocular regenerative medicine.

Background: Chronic wound healing is a complex clinical challenge, particularly due to microbial colonization and the deactivation of repair cells. This study presents an innovative strategy involving the combination of micron-scale chitosan fibers with exosomes, aiming to develop a new type of dressing with multiple functionalities, including dynamic exudate management, antimicrobial properties, angiogenesis promotion, and tissue repair. The goal is to offer a cost-effective and clinically translatable treatment solution for infected wounds.

Methods: Chitosan (CS) fibers were prepared using a wet-spinning technique and subsequently modified through N-succinylation (NCS), followed by needle-punching to construct CS/NCS blended nonwoven fabrics. The physicochemical properties, water absorption/retention, and mechanical behavior of the materials were characterized using scanning electron microscopy (SEM), Fourier-transform infrared (FTIR) spectroscopy, and X-ray diffraction (XRD). Antibacterial and hemostatic performance were also evaluated. Mesenchymal stem cell-derived exosomes (MSC-EXO) were loaded onto the modified chitosan fibers via electrostatic assembly, forming the CS/NCS-EXO composite dressing, which was further tested in a rat model for infected wound repair.

Results: Compared to pure CS fibers and NCS fibers, the CS/NCS material demonstrated superior mechanical properties and moisture retention capacity in a wet state. Antibacterial assays showed that the CS/NCS material exhibited significantly enhanced antimicrobial activity against Staphylococcus aureus and Escherichia coli. Hemostatic experiments revealed that the CS/NCS group significantly shortened bleeding time and reduced blood loss. In the infected skin defect repair experiment, the CS/NCS-EXO group significantly accelerated wound healing, demonstrating the most prominent tissue repair effect, accompanied by abundant angiogenesis as confirmed by immunohistochemical staining.

Conclusion: This study successfully developed a chitosan fiber-based exosome composite dressing system, which effectively coordinates infection control and tissue regeneration through a triple mechanism of "structural water-locking, mechanical adaptation, and bioactive synergy." This material provides a scalable solution for chronic wound management and shows promising clinical application prospects.

Background: Silicosis is a progressive lung fibrosis lacking effective treatment. Mesenchymal stem cells (MSCs) show antifibrotic potential, but their survival is impaired by the early inflammatory microenvironment. The therapeutic value of repeated MSC administration remains unclear.

Methods: A murine silicosis model was analyzed by single-cell RNA sequencing, bronchoalveolar lavage fluid (BALF) cytokine assays, and human Bone Marrow-Derived Mesenchymal Stem Cells (hBMSCs) transcriptomics after BALF exposure. Mice received either single or repeated intratracheal hBMSCs doses. Cell retention, lung function, imaging, histology, and fibrosis markers were assessed. The role of ZC3H4 in macrophage activation was examined by in vivo expression profiling, in vitro knockdown, and functional assays.

Results: Early silica exposure triggered strong M1 inflammation, high BALF cytokines, and hBMSCs senescence signatures. Repeated hBMSCs dosing improved cell persistence, reduced fibrosis on imaging and histology, enhanced lung function, and decreased collagen deposition compared with a single dose. Mechanistically, MSC therapy suppressed macrophage ZC3H4 expression, while ZC3H4 knockdown reduced macrophage activation and fibroblast migration.

Conclusions: Repeated hBMSCs administration enhances therapeutic efficacy in silicosis by improving cell persistence and attenuating fibrosis, partly through ZC3H4-mediated regulation of macrophages.

Background: The human endometrium is a regenerative tissue relying on stem/progenitor cells. Endometrial mesenchymal stem cells (eMSCs) are typically enriched using perivascular markers like CD140b and CD146. However, the identity of more primitive and quiescent eMSC subpopulations remains unclear.

Methods: We performed single-cell RNA sequencing (scRNA-seq) on cultured CD140b⁺CD146⁺ eMSCs and integrated this with published scRNA-seq data of primary human endometrial cells. We identified a LEPR⁺ subpopulation and analyzed its characteristics through in vitro assays, flow cytometry, immunostaining, and bioinformatic tools including cell-cell interaction analysis and pseudotime trajectory inference.

Results: A LEPR⁺ eMSC subpopulation was found to reside at the root of the differentiation trajectory and showed high expression of Notch receptors. These cells exhibited quiescent features, resided predominantly in the G0 phase, and demonstrated superior clonogenic and self-renewal capacity compared to LEPR⁻ eMSCs and bulk eMSCs. Notch signaling, particularly via JAG1 and DLL1, was implicated in maintaining the LEPR⁺ phenotype and quiescence.

Conclusions: LEPR⁺ eMSCs represent a primitive, quiescent subset of human endometrial stem cells. Notch signaling maintains their stemness and quiescence, suggesting therapeutic relevance for endometrial regeneration.

Background: Peripheral nerve injuries are associated with significant morbidity, particularly when primary surgical repair is delayed or impossible due to extensive nerve gaps. While advances in biomedical engineering have led to commercially available nerve guidance conduits tailored for such injuries, rates of sensory and motor recovery remain suboptimal following neurotmesis with gap defects beyond 3 cm. Cell therapy represents a promising treatment strategy to heal the injured peripheral nervous system, thought to promote tissue regeneration and enhance endogenous mechanisms of nerve repair to restore functionality. In this study, we explore the potential utility and efficacy of subcutaneous adipose tissue-derived neural stem cells in a nerve transection injury model.

Methods: Plp1-EGFP mice, which express GFP in Schwann cells, underwent surgical excision of a 5 mm segment of the left sciatic nerve. Nerves were then immediately repaired using silicone nerve guidance conduits with a residual 5 mm defect between nerve stumps. Conduits were loaded with cell culture media alone or with subcutaneous adipose tissue-derived neural stem cells harvested from Wnt1-tdTomato neural crest reporter mice, the latter enabling cell tracing post-transplantation.

Results: Subcutaneous adipose tissue-derived neural stem cells persisted through postoperative day 56 and contributed structurally to the reformed sciatic nerve. Integration between Wnt1-tdTomato neural stem cells and endogenous Plp1-EGFP Schwann cells occurred at the distal and proximal transection margins. Furthermore, neural stem cells predominantly differentiated into Schwann-like cells following transplantation, aiding in myelination of the reformed nerve, but not undesirable cell types such as neurons. Gait testing indicated that adipose-derived neural stem cells significantly improved hindlimb motor recovery compared to conduit repair alone by postoperative day 56.

Conclusions: Using cell tracer models, we confirm that adipose-derived neural stem cells can be therapeutically delivered to injured peripheral nerves, integrate with recipient axons and Schwann cells, and differentiate into myelinating Schwann-like cells to enhance motor recovery. These findings indicate that subcutaneous adipose tissue-derived neural stem cells could fill a critical gap in the treatment of peripheral nerve injuries, representing a readily available, autologous source of regenerative cells to optimize functional recovery after injury.

Background: Hereditary persistence of Fetal Hemoglobin (HPFH) is a benign condition known to mitigate symptoms in individuals with co-inherited β-hemoglobinopathies, such as β-thalassemia (BT) and sickle cell disease (SCD), through the reactivation of fetal hemoglobin (HbF). HPFH typically arises from deletions of varying sizes affecting the β-globin gene cluster or point mutations in the promoters of the γ-globin genes. While the therapeutic benefits of point mutations have been extensively studied, the potential of deletional forms of HPFH remains underexplored in preclinical settings.

Method: In this study, we generated benign deletional HPFH3 genotype in SCD and BT patient-derived HSPCs using CRISPR/Cas9 and showed that therapeutically relevant levels of HbF reactivation result in the alleviation of the pathological phenotypes.

Results: In edited cells derived from SCD patients, we observed reduced sickling and oxidative stress, while in edited from BT cells, restoration of the α-globin/β-globin ratio improved erythroid lineage maturation and reduced ROS levels. Importantly, HPFH3-edited HSPCs retained their genome integrity and showed no detrimental effect on their regeneration or differentiation into erythroid, myeloid, T, and B cell lineages in immunodeficient NBSGW mice post-xenotransplantation. Additionally, we showed a reduced interaction between the LCR and HBB, suggesting that the HPFH3 deletion specifically promoted LCR interactions with HBG1/2, likely due to the absence of the HBB locus.

Conclusions: Collectively, our preclinical findings suggest that the generation of the HPFH3 genotype has the potential to significantly enhance HbF levels, offering a promising universal therapeutic strategy for treating both SCD and β-thalassemia.

Background: Intestinal epithelial stem cells (SCs) and their transit-amplifying (TA) progeny are critical for mucosal repair and regeneration. However, their behaviour under chronic inflammatory conditions, such as those observed in Inflammatory Bowel Disease (IBD), remains incompletely understood.

Methods: We investigated the impact of chronic inflammation on intestinal stem/progenitor cells by integrating bulk RNA sequencing from the largest IBD biopsy cohort to date with single-cell transcriptomic analysis and experimental assays using patient-derived intestinal organoids.

Results: Active inflammation was associated with a reduction in canonical LGR5⁺ intestinal stem cells and a concurrent expansion of OLFM4⁺ populations, consistent with an inflammation-induced epithelial repair program. Notably, SC/TA cells from both inflamed and non-inflamed IBD tissues exhibited persistent transcriptional changes that were distinct from those in healthy controls. Single-cell analysis identified transcriptionally heterogeneous SC/TA subpopulations, including a previously uncharacterized inflammation-associated cluster enriched in immune signalling pathways. Pseudotime trajectory analysis demonstrated a shift in differentiation toward deep crypt secretory (Paneth-like) cell lineages under inflammatory conditions.

Conclusions: Chronic intestinal inflammation reshapes the epithelial stem and progenitor cell compartment, promoting altered differentiation and the emergence of immune-responsive epithelial states. These findings highlight the plasticity of the human intestinal epithelium in IBD and point to new avenues for therapeutic strategies aimed at maintaining epithelial integrity during chronic inflammation.