Stem Cell Research & Therapy最新文献

Background: Cell therapy has been proposed as a promising treatment for neurological recovery in patients with stroke. However, a strategy to enhance its efficacy is needed, as its clinical benefits have not yet been demonstrated in clinical trials. This study evaluated the efficacy of combination therapy using allogeneic umbilical cord blood (UCB), a relatively safe therapeutic cell source, and recombinant human erythropoietin (rhEPO) in patients with subacute stroke.

Methods: In this double-blind, randomised controlled trial, we enrolled patients with subacute stroke one to nine months after stroke onset. The patients were divided into three groups: UCB + EPO, UCB, and control. Immune compatibility-matched UCB was intravenously infused once, and rhEPO was administered five times. Safety was evaluated according to the Common Terminology Criteria for Adverse Events (version 5.0), while efficacy was assessed based on changes in activities of daily living, motor and cognitive functions, brain imaging findings, and electroencephalography performed at six months after baseline.

Results: A total of fifteen patients (59.0 ± 10.9 years) were included, with consisting of five patients in each group with comparable demographic data and functional parameters at baseline. Adverse events did not indicate any harmful effects of UCB or rhEPO. After all patients completed the final functional evaluation the UCB + EPO group showed significantly better outcomes than the control group in terms of the total Functional Independent Measure (FIM) (Δ15.00[12.50, 24.50] vs. Δ0.00[-13.00, 3.00], P = 0.009), FIM motor subscale (Δ14.00[10.00, 18.50] vs. Δ13.00[0.50, 3.50], P = 0.009), and Geriatric Depression Scale (Δ-3.00[-5.00, -2.00] vs. Δ6.00 [-1.00, 18.50], P = 0.016) scores. The UCB group showed a marginally non-significant improvement over the control group, without statistical differences in most outcome measures. The brain imaging findings also supported the functional recovery-related effects of UCB therapy.

Conclusion: In conclusion, rhEPO can enhance the efficacy of UCB cells in patients with subacute stroke, without causing harmful effects. This exploratory finding may provide evidence for the potential use of UCB + EPO combination therapy for neurological recovery following stroke. Trial registration URL: https://clinicaltrials.gov/ct2/show/NCT04013646 .

Background: Acute lung injury/acute respiratory distress syndrome (ALI/ARDS) remains a critical respiratory condition with limited effective treatments.

Methods: This study investigated whether mesenchymal stem cells (MSCs) preconditioned with supernatant from hypoxia-cultured telocytes (TCs) could enhance therapeutic efficacy in ALI through regulatory T cell (Treg) modulation.

Results: MSCs preconditioned with 5% hypoxic TC supernatant demonstrated superior efficacy in ameliorating LPS-induced lung injury compared to conventional MSCs or TC monotherapy, as evidenced by preserved alveolar architecture, reduced inflammatory infiltration, and decreased pro-inflammatory cytokines. Mechanistically, these preconditioned MSCs significantly enhanced Treg recruitment to injured lung tissues and improved their immunosuppressive function through the CXCL5/6-CXCR1 axis, an effect that was substantially attenuated upon siRNA-mediated disruption of this pathway, and was further corroborated in a humanized ALI mouse model where preconditioned-MSC treatment improved survival, reduced lung injury severity, and enhanced Treg recruitment and function in a CXCL5/6 signaling-dependent manner.

Conclusions: These findings reveal a novel mechanism by which hypoxic TC supernatant enhances MSC therapeutic efficacy in ALI through the CXCL5/6-CXCR1 axis, providing a promising strategy for optimizing cellular therapy in inflammatory pulmonary disorders.

Background: The resolution of inflammation is an active process triggered in the acute phase of inflammation and mainly directed by specialized pro-resolving mediators (SPMs). Modulating the inflammatory response in favor of resolution is a therapeutic strategy of enormous interest and value. Previous studies have shown that human adipose mesenchymal stem cells (MSCs) possess the ability to shorten the acute inflammatory response by hinting an early cyclooxygenase (COX)-2 induction. We studied the potential of MSC to accelerate the resolution of inflammation through the direct promotion of the local synthesis of SPMs, and the role of prostaglandin (PG) E₂ release in this process in a model of experimental acute arthritis.

Methods: Gouty arthritis was induced in male New Zealand rabbits via intra-articular injection of monosodium urate (MSU) crystals. Human adipose-derived MSCs were administered systemically in a single dose. Synovial membrane levels of SPMs were measured by liquid chromatography-tandem mass spectrometry, PGE₂ and genes related to the pro-resolving and anti-inflammatory pathways were assessed. Employing THP-1-derived macrophages and human adipose-derived MSC co-cultures stimulated with MSU crystals, we elucidated the mechanisms associated to the induction of resolution by MSCs.

Results: MSC treatment enhanced the local release of a broad range of SPM precursors and active mediators in the synovial membrane of rabbits with gouty arthritis. This release occurred simultaneously with an early increase in PGE₂ levels, and an upregulation of COX-2 and the PGE₂ receptor EP4. Additionally, an early anti-inflammatory gene response was observed, characterized by increased expression of IL-10, indoleamine 2,3-dioxygenase-1 (IDO-1), and Formyl Peptide Receptor2 (FPR2). In cell culture experiments, we confirmed that MSCs are responsible for the release of pro-resolving and anti-inflammatory mediators, promoting macrophage efferocytosis and polarization towards a pro-resolving and anti-inflammatory M2 phenotype in a COX-2- and FPR2-dependent manner.

Conclusions: MSCs exerted a pro-resolving effect on the synovial membrane in gouty arthritis. This therapeutic action may be driven by an early superinduction of local PGE₂ synthesis and the promotion of a pro-resolving and anti-inflammatory M2 macrophage phenotype via COX-2 signaling and involving FPR2.

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.