Stem Cell Research & Therapy最新文献

Background: Decidualization, the process of endometrial stem/stromal cell (EnSCs) differentiation, is essential for embryo implantation and pregnancy maintenance. Menstrual blood-derived stem/stromal cells (MenSCs), although often considered as surrogates of EnSCs, represent a distinct population. Pharmacologic modulation of cellular senescence using senomorphics has emerged as a promising strategy in reproductive medicine. This study investigates the decidualization capacity of EnSCs and MenSCs and evaluates how senomorphic agents influence their senescence, metabolic profile, and inflammatory response.

Methods: Primary EnSCs and MenSCs were isolated, characterized, and subjected to in vitro decidualization using standardized protocols. Cells were classified as well-decidualized (WD) or poorly-decidualized (PD) based on the extent of decidualization. Six senomorphic compounds were applied before and during decidualization. Senescence-associated β-galactosidase activity, IL-6 secretion, glycolytic intermediates, and global metabolomic changes were assessed before and after treatment with senomorphics.

Results: MenSCs exhibited accelerated but limited and prolonged decidualization capacity compared to EnSCs. Metabolic reprogramming in EnSCs at day 6 resembled that of MenSCs at day 3. Decidualization induced differential changes in glycolysis-related metabolites, senescence markers, and IL-6, especially in PD cells. Treatment with six senomorphics modulated these effects in a context-dependent manner. Exposure during decidualization increased senescence in both WD and PD sources, whereas pretreatment increased senescence in WD EnSCs but decreased it in PD EnSCs. Notably, senomorphics shifted the metabolomic profile of PD EnSCs toward a WD-like state.

Conclusions: EnSCs and MenSCs differ in decidualization dynamics, metabolism, and response to senomorphic modulation. Senomorphics may be strategically employed to reduce senescence in patients with impaired endometrial decidualization, offering therapeutic potential in reproductive pharmacology.

Background: Intrauterine adhesion (IUA) is a common gynecological disease that contributes to infertility. Decreased endometrial angiogenesis and uterine ischemia are major therapeutic challenges for IUA and cannot be addressed by current treatment strategies. Human endometrial mesenchymal stem cells (H-EMSCs) and macrophages (mø) are both important cell types that reside within the endometrial tissue and participate in its repair and regeneration. However, how to harness the endometrial tissue repair potential of H-EMSCs and mø simultaneously in a co-delivery system and whether there are significant biochemical cross-talks between the two cell types so that they can regulate each other to specifically boost endometrial tissue angiogenesis remains to be explored.

Methods: This study developed a H-EMSCs-mø co-delivery system using an electrospun polycaprolactone-hyaluronic acid (PCL-HA) membrane and established a rat endometrial damage model. The effects of the co-delivery system on endometrial tissue repair (endometrium thickness, endometrial glands number) and angiogenesis were investigated. The mechanisms underlying the enhanced endometrial tissue angiogenesis of the H-EMSCs-mø co-delivery system were also delineated. All data were analyzed using analysis of variance with Tukey's test for pair-wise comparisons or an independent samples t-test where appropriate.

Results: In this study, it was found that a H-EMSCs and mø co-delivery system developed with a PCL-HA electrospun membrane carrier (PCL-HA/H-E/mø) significantly increased the endometrium thickness and restored the number of endometrial glands at day 7 and 14 in the endometrial damage model vs. the NR (normal repair) and PCL-HA alone groups. Further, PCL-HA/H-E/mø enhanced more CD31 gene and protein expression, indicating great potential for angiogenesis to occur at day 7 and 14 post-implantation, when compared with PCL-HA/H-E, NR or PCL-HA alone. It was also proved to demonstrated that elevated VEGF production was one of the potential factors that contributed to the enhanced angiogenesis of the co-delivery patch system.

Conclusions: This study provided significant insights into the use of co-delivered H-EMSCs and mø, on a PCL-HA hybrid electrospun membrane, for effectively inducing endometrial angiogenesis and repair to enhance IUA treatment outcomes.

Background: The abnormal immune response mediated by CD4⁺T cells is a key factor in Immune thrombocytopenia(ITP) progression. While Ningxue Shengban Decoction (NXSBD) is an effective therapeutic, its underlying mechanism and targets remain obscure.

Aim: This study aims to clarify the role of exosomal miR-199a-5p derived from bone marrow mesenchymal stem cells (BMSCs) in immune homeostasis, and to explore the therapeutic effects of exosomes from BMSCs(BMSCs-Exo) pretreated with NXSBD containing serum on ITP.

Method: We co-cultured CD4⁺T cells with BMSCs or pre-treated BMSCs-Exo. The proliferation and differentiation of CD4⁺T cells were then assessed using CFSE staining and flow cytometry (FCM). Additionally, an active ITP murine model was employed to assess the therapeutic efficacy of pre-treated BMSCs-Exo. Platelet counts were measured and organ indices were calculated. Serum autoantibody levels were measured by FCM and ELISA, changes in CD4⁺T cells subsets in the spleen were analyzed by FCM, megakaryocyte number and morphology in bone marrow tissues were examined by H&E staining, and key cytokine levels in mouse serum were quantified by ELISA.

Results: Our results indicate that the immunomodulatory effect of BMSCs-Exo on CD4⁺T cells is mediated by miR-199a-5p, and that NXSBD containing serum enhances this effect by increasing miR-199a-5p levels. In an active ITP murine model, BMSCs-Exo treatment significantly ameliorated the pathological features of ITP, as evidenced by increased peripheral platelet counts, reduced spleen and thymus indices, and decreased levels of autoantibodies. Immunophenotypic analysis revealed that an increased percentage of splenic Treg and Th2 cells, and a decreased percentage of Th17 and Th1 cells, were observed after BMSCs-Exo treatment. Additionally, BMSCs-Exo enhanced the production of mature megakaryocytes. Following BMSCs-Exo treatment, the levels of pro-inflammatory cytokines were sharply lowered, whereas anti-inflammatory cytokine levels were markedly elevated. BMSCs-Exo pretreated with NXSBD containing serum exert superior therapeutic efficacy compared with those derived from untreated BMSCs.

Conclusion: In conclusion, our findings suggest that the therapeutic effect of NXSBD containing serum in ITP mice may be attributed to its upregulation of miR-199a-5p in BMSCs-Exo, which contributes to modulating the immune balance of CD4⁺T cells.

Osteoarthritis (OA) is a complex disease associated with genetic, biological, and mechanical risk factors that act, in part, to alter chondrocyte homeostasis. Our recent exome sequencing studies identified a damaging genetic variant in COL6A3, a monomeric unit of collagen type VI and a distinguishing component of the pericellular matrix (PCM) of articular cartilage, a transducer of mechanical and biochemical signals for the chondrocyte. To study the effect of this genetic variant, human induced pluripotent stem cell (hiPSC)-derived chondrocytes, genetically edited to harbor the COL6A3 mutation, were used as an in vitro model to investigate chondrocyte mechanobiology and pathobiology. The COL6A3 variant resulted in lower PCM elastic modulus and reduced expression of key matrix proteins, suggesting altered PCM structural composition and mechanical properties. Functional analyses revealed altered mechanotransduction, characterized by heightened osmotically-induced calcium signaling, consistent with reduced PCM modulus, and reduced anabolic response to TRPV4 activation, both at the transcriptional level and in matrix biosynthesis. RNA-sequencing identified dysregulated pathways and aberrant TRPV4 signaling in mutant chondrocytes following mechanical loading. The presence of the COL6A3 variant also resulted in disrupted circadian rhythms, with increased BMAL1 expression and a significant phase shift, suggesting that PCM properties influence the circadian clock. Finally, COL6A3 mutant chondrocytes exhibited an exacerbated catabolic response to interleukin-1, an inflammatory cytokine implicated in OA. Our study demonstrates the utility of human iPSCs for studying the pathophysiology of specific OA risk alleles. These findings highlight the impact of the COL6A3 variant on chondrocyte physiology and support targeting mechanotransduction signaling pathways as a potential strategy for OA intervention.

Background: Vascular dysfunction caused by urethral injury often leads to delayed repair. Exosome from stem cell showed promise in tissue regeneration. But human induced pluripotent stem cell-derived exosomes (hiPSC-Exo) has not been reported the angiogenesis ability in urethral injury repair and their potential mechanisms.

Method: The exosome was extracted from hiPSCs. The in vivo and in vitro experiments were performed to investigate the effects of hiPSC-Exo on angiogenesis. The miRNA-seq bioinformatics, luciferase assay and related functional experiments were performed to determine potential mechanism.

Result: Exosome was extracted from hiPSCs by ultracentrifugation. Compared to the control group, hiPSC-Exo significantly promoted blood flow supply to the ischemic lower limbs of mice and rat model of urethral defects to accelerate injury repair. We found that hiPSC-Exo significantly enhanced the proliferation, migration, and tube formation ability of endothelial cells in vitro. MiRNA-seq analysis and experiments verified that miR-103a-3p was highly expressed in HUVEC treated with hiPSC-Exo and significantly enhanced the proliferation, migration, invasion, and angiogenesis effects. TGFBR3 was identified as a direct target of miR-103a-3p through bioinformatics, qPCR, and dual luciferase assays. Overexpression of TGFBR3 leaded to reduced proliferative, migrative and angiogenesis ability of HUVEC, but silence could promote HUVEC function. TGFBR3 could decrease VEGF expression and phosphorylation-based activation of FAK.

Conclusion: This study indicated that hiPSC-Exo played a crucial role in promoting angiogenesis to accelerate urethral injury repair through the action of miR-103a-3p in exosome on the TGFBR3/VEGF/FAK signaling pathway in endothelial cells. This provides a new treatment strategy for hiPSC-Exo in the clinical treatment of urethral injury healing and elucidates its unique mechanism of action.

Background: Hematopoietic stem and progenitor cells (HSPCs) are crucial for blood production and regeneration. While their function is known to be regulated by diverse physical cues, the impact of pervasive radiofrequency electromagnetic fields (RF-EMF), particularly through non-thermal radiofrequency radiation (RFR) mechanisms, remains poorly understood.

Methods: We conducted colony-forming unit (CFU) assay in vitro and competitive transplantation assay in vivo to evaluate whether RFR influences hematopoiesis reconstitution capacity. Subsequently, the effects of RFR preconditioning on hematopoietic injury induced by ionizing radiation in mice were assessed by continuously monitoring the peripheral blood, HSPCs number, and colony-forming units. The influence of RFR on radioprotection unit frequency was evaluated using multiple gradients, non-competitive mouse transplantation models. Seahorse XF assays were employed to characterize cellular energy metabolic status, while specific fluorescent probes were utilized to detect calcium ion (Ca²⁺) levels in distinct cellular compartments using flow cytometry. Transcriptomic profiling was used to uncover the underlying mechanisms. HSPCs were pretreated with plasma membrane Ca²⁺-ATPase (PMCA) inhibitor prior to RFR exposure, and Seahorse assays along with CFU assay and competitive transplantation assay were performed to compare whether PMCA inhibition could abrogate RFR-induced HSPCs function change. To investigate the mechanism by which RFR enhanced PMCA activity inducing Ca²⁺ efflux, we performed fluorescence recovery after photobleaching (FRAP) assays to detect membrane fluidity.

Results: Non-thermal 2856 MHz RFR enhanced HSPCs colony formation activity and reconstitution capacity, without compromising the multilineage differentiation homeostasis. RFR preconditioning accelerated hematopoietic recovery following ionizing radiation and increased radioprotection unit frequency. Mechanistically, RFR increased plasma membrane fluidity which potentiates PMCA activity, resulting in elevated Ca²⁺ efflux and reduced intracellular Ca²⁺ levels. These cellular alterations ultimately contributed to maintaining HSPCs in a low metabolic state, and consequently improving their functional capacity. Pharmacological inhibition of PMCA abolished both the functional enhancement and metabolic suppression.

Conclusion: Our results provided the first evidence that non-thermal RFR can improve HSPCs function. The central mechanism involved RFR-induced plasma membrane fluidity, activation of PMCA, thus accelerating Ca²⁺ efflux and maintaining HSPCs in a metabolically quiescent state. This work provided transformative insights into electromagnetic field biology and potential transplantation strategies for radiation-induced hematopoietic injury.

Background: Mitochondrial transplantation (Mito-T) is a novel therapeutic strategy for ischaemic cardiovascular diseases. This study aimed to test the efficacy of human umbilical mesenchymal stem cell-derived mitochondrial transplantation (Mito-T) on preeclampsia (PE).

Methods: PE was induced in Sprague-Dawley pregnant rats by infusing angiotensin II (Ang II) starting on gestation day 8 (GD 8). Mito-T (100 μg/μl) was injected via the jugular vein on GD 14.

Results: On GD 20, PE rats exhibited high blood pressure, kidney and placental vascular abnormalities, reduced placental and foetal weights, foetal crown-rump lengths. Mito-T was predominantly distributed in the kidneys, uterus, and placenta of PE rats. Mito-T reversed clinical manifestations of PE, restored placental vascular abnormalities, and reduced serum sFLT-1 levels and the sFLT-1/PlGF ratio. In placental mitochondria, Mito-T increased protein levels of complexes (I‒V), improved mitochondrial membrane potential, ATP synthase, citrate synthase activities, and biogenesis markers (PGC-1α, TFAM, and NRF1), and reduced reactive oxygen species production. Mito-T increased mitochondrial fusion proteins (OPA1, MFN1, and MFN2) in the placenta, whereas fission (DRP1 and FIS1) and mitophagy (PINK, BNIP3, BNIP3L, and FUNDC1) proteins were reduced. In placental tissue, primary trophoblast cells, and the Bewo cell line, Mito-T reduced the mRNA and protein levels of sFLT-1 and attenuated the calcineurin-NFAT pathways elevated by PE or Ang II.

Conclusions: This study demonstrates that Mito-T reverses the pathological phenotypes of PE rats by improving placental mitochondrial activity and suppressing trophoblast-derived sFLT-1 production. These findings provide proof-of-concept evidence that Mito-T could serve as a potential therapeutic strategy for reducing maternal and foetal risks in patients with PE.

Background: Cardiac fibrosis represents a significant health burden, with endothelial dysfunction and damaged perivascular microenvironment increasingly recognized as key contributors to fibrotic remodeling. The urokinase plasminogen activator receptor (uPAR), a critical component of the urokinase system, plays a pivotal role in vascular remodeling and fibrosis. While prior evidence indicates that uPAR deficiency leads to microvascular dysfunction and perivascular fibrosis, the underlying mechanisms remain poorly defined. This study investigates how uPAR deficiency contributes to fibrotic remodeling of the cardiac perivascular-like microenvironment.

Methods: Single-cell RNA sequencing data analysis and immunofluorescence staining on mouse heart cryosections were performed to characterize uPAR expression within the cardiac perivascular microenvironment. To model this microenvironment in vitro, cardiospheres (CSs) were generated from non-myocyte cardiac cells of wild-type and uPAR-knockout mice. CRISPR/Cas9-generated Plaur knockout (KO) 3T3 fibroblasts (FBs) were employed as model stromal cells. Pro-fibrotic activation of FBs was induced by TGFβ1 treatment. Comparative analyses of extracellular matrix (ECM) deposition, fibrotic cell transformation, and comprehensive secretome profiling was conducted using western blotting.

Results: Our findings demonstrated that uPAR was expressed by endothelial cells (ECs) and FBs within the cardiac perivascular microenvironment. uPAR deficiency exacerbated profibrotic stimuli in CSs, including elevated active TGFβ1, impaired integrin functions, and altered cell secretome. These alterations collectively disrupt critical cell-cell and cell-matrix interactions, leading to increased ECM deposition, EC loss and decreased cell viability. Using Plaur KO FBs, we demonstrated that uPAR deficiency amplified TGFβ1-mediated Akt signaling pathway and ECM deposition.

Conclusions: Our study reveals that uPAR loss drives fibrotic remodeling of the cardiac perivascular-like microenvironment and exacerbates TGFβ1-mediated effects, highlighting its potential as a therapeutic target for cardiac fibrosis.