Stem Cell Research & Therapy最新文献

Background: Renal ischemia-reperfusion injury (RIRI) refers to kidney damage following blood flow restoration, with limited effective treatments available. Bone mesenchymal stem cell (BMSC) derived exosomes exhibit therapeutic potential via targeted molecular delivery, though limited by isolation challenges and transient retention, while curcumin demonstrates multi-organ protective capacities in RIRI.

Methods: In vivo, RIRI mice received tail vein injections of BMSC exosomes (exo) or curcumin preconditioned BMSC exosomes (cur-exo). Biodistribution was tracked via bioluminescence/immunofluorescence, while therapeutic efficacy was evaluated through renal function parameters, histopathology, and ferroptosis biomarkers. In vitro, ferroptosis-induced renal tubular epithelial cells were treated with exo and cur-exo, with subsequent quantification of Fe²⁺, lipid peroxidation, glutathione, mitochondrial ultrastructure, ROS levels, and ferroptosis-related protein/mRNA expression. Mechanistic studies integrated transcriptomics, siRNA/overexpression systems, ChIP, dual-luciferase assays, SPR, Co-IP and bioinformatics to delineate anti-ferroptosis pathways of cur-exo and the effect of curcumin on miR-16-5p.

Results: Curcumin preconditioning can enhance the targeted delivery capability of BMSC exosomes to injured kidneys and improves the restoration of renal function and tissue damage in mice with ischemia-reperfusion injury by inhibiting ferroptosis. In vitro, TCMK-1 cells can take up both exo and cur-exo, with cur-exo significantly enhancing the survival rate of TCMK-1 cells induced by ferroptosis compared to exo. This is achieved by downregulating lipid peroxidation levels, improving iron overload and ROS accumulation, and restoring mitochondrial structure to exert anti-ferroptosis effects. Mechanistically, curcumin increases the expression of miR-16-5p in cur-exo by regulating the activity of CYP1B1, and cur-exo inhibits the translation of Smad3 by delivering miR-16-5p that targets the 3'UTR of Smad3, leading to the downregulation of myoglobin (Mb) transcriptional activity and thereby antagonizing ferroptosis in TCMK-1 cells.

Conclusion: Our research indicates that curcumin preconditioned BMSC exosomes can exert a therapeutic effect on RIRI by inhibiting cellular ferroptosis. The primary mechanism behind this effect involves curcumin increasing the expression of miR-16-5p by modulating CYP1B1 activity, and cur-exo promoting the alleviation of ferroptosis in TCMK-1 cells through the miR-16-5p/Smad3/Mb axis. This study provides a new strategy for enhancing the biological functions of exosomes and presents new targets and ideas for the treatment of RIRI.

The convergence of CRISPR genome editing, patient-derived organoids, and induced pluripotent stem cells (iPSCs) has reshaped in vitro disease modeling by enabling mechanistic investigations of human pathophysiology within genetically matched, tissue-relevant systems. Together, these technologies provide a synergistic platform for precise manipulation of disease-associated variants and support the generation of isogenic organoid models that reproduce key phenotypic and functional hallmarks across cancer, neurodegenerative, inflammatory, and monogenic disorders. In this review, we highlight how diverse CRISPR modalities-including knock-out, knock-in, CRISPRa/i, and genome-scale screening-have been applied to dissect gene function, model disease progression, and guide therapeutic development using iPSC- and organoid-based systems. We further discuss the application of these platforms in genotype- and phenotype-driven precision medicine, enabling patient stratification, drug-response prediction, and individualized treatment design. We illustrate these convergent applications with representative case studies spanning mechanistic research and early clinical translation. By combining the scalability of genome engineering with the physiological fidelity of organoids, CRISPR-integrated platforms are redefining the frontiers of experimental medicine. These approaches accelerate the discovery of disease mechanisms and actionable therapeutic targets while establishing individualized clinical strategies for complex human diseases. Collectively, they position CRISPR-enabled organoid systems as a foundational infrastructure that bridges genome editing to individualized therapy and supports next-generation precision medicine.

Alopecia Areata (AA) is a chronic inflammatory disorder characterized by non-scarring, patchy hair loss that may progress to the entire scalp (alopecia totalis) or body (alopecia universalis), significantly impairing patients' quality of life and psychological health. Although the exact pathogenesis of AA remains unclear, current evidence suggests that the breakdown of hair follicle immune privilege (IP) and subsequent autoimmune-mediated follicular attack play a pivotal role. Conventional therapeutic modalities, including corticosteroid and Janus kinase (JAK) inhibitors, are often limited by suboptimal efficacy in severe cases and high relapse rates following treatment cessation. In recent years, stem cell-based therapy has emerged as a novel treatment for AA, showing therapeutic potential through multiple mechanisms. Preliminary clinical trials have indicated significant efficacy in promoting hair regrowth among AA patients. However, comprehensive evaluation of long-term safety and therapeutic efficacy remains imperative. This review article aims to give a comprehensive overview of the recent advances in stem cell-based therapies for AA and explore their underlying mechanisms and clinical application prospects, hoping to provide a framework and reference for future research and clinical practice.

Mesenchymal stromal cells (MSCs) are widely recognized for their immunomodulatory properties, which underpin their therapeutic potential in inflammatory and immune-mediated diseases. Although MSC therapies have consistently proven safe, clinical efficacy remains inconclusive, maybe due to incomplete understanding of MSC interactions with the immune environment. This review evaluates current trends in MSC immunomodulation research, based on 318 studies published since 2019 until medio 2024. The most frequently used assays included characterization, proliferation, and polarization, employing methods such as flow cytometry, enzyme-linked immunosorbent assays and colorimetric assays, and polymerase chain reaction. Many studies incorporated strategies for priming of MSCs or included immune cells, most commonly peripheral blood mononuclear cells, T cells, and macrophages. We identify key sources of variability and propose a minimum reporting checklist including MSC source, priming conditions, assay design, and immune cell characteristics. We further recommend implementation of multi-assay workflows combining phenotypic characterization with at least one functional assay. These measures may improve transparency, comparability across studies, and guide robust assay design.

Background: Protein tyrosine phosphatases (PTPs) play key roles in β-cell function and diabetes development. PTPN2 is a candidate gene for type 1 diabetes (T1D) that negatively regulates JAK/STAT signalling. However, the impact of PTPN2 deficiency on the differentiation and functionality of human stem cell-derived somatic metabolic cells remains unclear.

Methods: PTPN2 expression in β cells from T1D organ donors and during the differentiation of human stem cell-derived islets (SC-islets) was evaluated using single-cell RNA-Sequencing (scRNA-Seq) datasets. We differentiated CRISPR-Cas12a genome-edited PTPN2-deficient H1 human embryonic stem cells (H1-hESCs) into SC-islets, and scRNA-Seq was performed. The maturation and functionality of PTPN2-deficient SC-islets were assessed by implantation under the kidney capsule of NOD-SCID mice.

Results: scRNA-Seq analysis showed that PTPN2 expression was increased in β cells from recently diagnosed T1D and decreased in long-standing T1D organ donors compared with controls. Conversely, we found that PTPN2 expression was decreased at the early stages of SC-islet differentiation and reconstituted at the later stages, suggesting a developmental dynamic. PTPN2 deficiency exacerbated interferon-induced inflammatory signalling in stem cells and differentiated somatic metabolic cells. Interestingly, PTPN2 deficiency increased hedgehog signalling and reduced SC-islet differentiation efficiency in vitro. In addition, PTPN2-knockout SC-islets exhibited reduced glycaemic control after implantation in vivo, mediated by reduced endocrine cell identity and enhanced interferon signalling.

Conclusions: Our study postulates a key role of PTPN2 in preserving β-cell function during inflammatory and metabolic stress in SC-islets.

Objective: Heterotopic ossification (HO) is a pathological condition characterized by dysregulated regenerative processes in skeletal muscle, resulting in the formation of mature bone within ectopic sites. Accumulating studies indicate that M2 macrophages facilitate endothelial-mesenchymal transition (EndMT), which contributes centrally to HO pathogenesis. This study explored the mechanism of M2 macrophage involvement in EndMT-mediated HO and analyzed the underlying molecular pathways.

Methods: Clinical samples of immature, mature, and autologous cancellous bones were collected to analyze macrophage infiltration and the cellular origin of HO. Using the GEO and GeneCards databases, MSX2 was identified as a candidate regulatory factor. An endothelial-macrophage co-culture system was established to investigate the specific molecular mechanisms by which macrophages affect EndMT by modulating MSX2 expression. Achilles tendon incision surgery was performed in C57BL/6 mice to simulate trauma-induced HO. Histological examination, immunohistochemistry, immunofluorescence, and ELISA were performed to verify the role of macrophages in influencing HO progression via MSX2. The STRING database was used to predict LEF1 as a downstream target gene of MSX2. Regulatory interactions between MSX2 and LEF1 were examined using dual-luciferase reporter assays, western blotting, and quantitative real-time PCR.

Results: Compared to autologous cancellous bone, there was an increase in M2 macrophage infiltration in HO tissues, accompanied by the transformation of endothelial cells at the injury site into mesenchymal stem cells. The degree of HO positively correlated with elevated levels of BMP-2, SP, Act A, TGF-β, OSM, and NT-3 in the serum. In vitro experiments demonstrated that under co-culture conditions, M2 macrophages induced mouse aortic endothelial cells (MAOECs) to exhibit elevated expression of mesenchymal markers (N-cadherin, vimentin) and mesenchymal stem cell markers (CD44, CD90), while reducing levels of endothelial markers (E-cadherin, occludin). Moreover, M2 macrophage-driven EndMT activation further promoted the upregulation of chondrogenic (Sox9, SP7) and osteogenic (OPN, OCN, Runx2) markers. However, MSX2 depletion rescued M2 macrophage-induced EndMT activation. In vivo, MSX2 inhibition or macrophage depletion reduced the osteogenic capacity in mice and decreased HO formation, whereas injection of a lentivirus overexpressing MSX2 promoted HO formation. Mechanistically, MSX2 directly binds to the LEF1 promoter to enhance its transcriptional activity, thereby activating Wnt signaling and maintaining EndMT.

Conclusions: Our findings suggest that M2 macrophages regulate EndMT-driven osteogenesis through MSX2/LEF1/Wnt axis, providing new insights into future therapeutic strategies for HO.

Background: As a chronic inflammatory disease characterized by periodontal tissue destruction, periodontitis has a pathogenesis that remains incompletely understood. This study aimed to investigate the expression of upregulated vanin-2 (VNN2) in exosomes derived from dental pulp stem cells (DPSCs) and its effects on the function of periodontal ligament stem cells (PDLSCs) and the progression of periodontitis.

Methods: A total of 35 patients with periodontitis and 35 healthy individuals were enrolled for gingival tissue sample collection. DPSC-EXO-VNN2 was cultured with a 3D system and isolated by ultracentrifugation. The samples were then subjected to nanoparticle tracking analysis (NTA), western blot and transmission electron microscopy (TEM). In vitro, PDLSCs were treated with DPSC- EXO-VNN2 and LPS, and the expression of IL-6 and TNF-α and their osteogenic potential were evaluated. Furthermore, in vivo, experimental periodontitis was induced in rats, which were then divided into two groups and treated with either DPSC-EXO-VNN2 or PBS. The maxillae were subsequently collected for histological staining and micro-CT analysis.

Results: VNN2 was upregulated in periodontitis according to dataset analysis, western blot and immunofluorescence (P < 0.05). Higher expression levels of VNN2 were associated with greater periodontal parameters PD and CAL (P < 0.05). The production of DPSC-EXO in the 3D culture system surpassed that in the 2D system. In vitro, PDLSCs internalized 3D-DPSC-EXO-VNN2, which increased LPS-induced IL-6 and TNF-α production while reducing osteogenic differentiation, as shown by decreased ALP activity and mineralization. In vivo, DPSC-EXO-VNN2 administration worsened alveolar bone loss, as micro-CT revealed a significantly lower bone volume fraction (P < 0.01) than did the control treatment.

Conclusion: This study reveals for the first time that DPSC-EXO-VNN2 participates in the progression of periodontitis by regulating the inflammatory response and osteogenic differentiation ability of PDLSCs. Future research may further explore the potential application of targeting VNN2 in treating periodontitis.

Background: Extracellular vesicles derived from human induced mesenchymal stromal cells (hiEVs) constitute a promising cell-free therapeutic option for osteoarthritis. To facilitate transition to the clinic we evaluated the therapeutic effects of hiEVs for osteoarthritis treatment. Specifically, we compared the efficacy of hiEVs collected from serum-containing and serum-free, PurStem (PS), media in an osteoarthritis mouse model.

Methods: hiEVs were administered via intra-articular injection in a destabilization of the medial meniscus (DMM) mouse model, with or without hydrogel to determine added value of localized application and controlled hiEV-release. Fluorescence imaging was used to monitor the retention of IR780-labeled hiEVs in the joint cavity. Therapeutic effects were evaluated by scoring of damage as well as expression of Mmp13 and Col2, catabolic and anabolic markers respectively, in joint tissues. Subchondral bone changes were assessed with Micro-CT.

Results: Fluorescence imaging confirmed that hiEVs remained localized at the injection site without systemic migration. HiEVs demonstrated significant protective effects against joint tissue degeneration in the osteoarthritis DMM mouse model as evidenced by reduced damage scores, decreased Mmp13 expression, and increased Col2 expression independent of the medium used for hiEV collection. The hydrogel alone also showed beneficial therapeutic effects, illustrated by reduced damage scores, increased Col2, and reduced Mmp13 expression. These effects, however, were notably smaller than those achieved with hiEV treatment. Micro-CT analysis further showed that hiEV treatment attenuated DMM-induced subchondral bone sclerosis as reflected by normalization of the bone volume fraction and trabecular structure.

Conclusions: Together, our findings demonstrate that hiEVs from xeno-free conditions effectively prevent cartilage degradation and promote its repair. This paves the way for future clinical translation of hiEV-based therapies as a safe, scalable, and effective approach to treat osteoarthritis.

Lung cancer is the first leading cause of cancer death worldwide. oxysterol-binding protein-like 2 (OSBPL2), is a lipid transport protein regulating cholesterol homeostasis. Here, we clarified the previously unreported role of OSBPL2 in lung cancer stemness properties. We observed that OSBPL2 reduced cholesterol content by HPLC-MS. It inhibited the accumulation of lipid droplets (LDs) in lung cancer. OSBPL2-mediated lipid transportation significantly suppressed tumor sphere formation, stemness markers expression and in vivo tumorigenesis and tumor metastasis. In clinical specimens, we also demonstrated that OSBPL2 repressed the expression of Lung cancer stem-like cells (LCSCs) markers-ALDH1A1, CD133 and Nanog. The level of OSBPL2 was negatively correlated with malignant of lung cancer, such as tumor stage progression and lymph node metastasis. Taken together, these findings illustrated that OSBPL2-mediated lipid transportation inhibited the stemness and aggressiveness of lung cancer cells. OSBPL2 was a potential therapeutic target to develop novel cancer-preventive compound.

Atopic dermatitis (AD) is a chronic inflammatory skin disorder characterized by disrupted epidermal barrier function, immune dysregulation, and persistent inflammation. Affecting both children and adults, AD significantly impairs quality of life due to its visible symptoms and associated psychosocial and economic burdens. Traditional treatments such as application of topical corticosteroids, calcineurin inhibitors, moisturizers and antibiotics, while managing symptoms, often fall short of providing long-term solutions and can lead to adverse effects over time. This review explores innovative approaches to AD management, focusing on the therapeutic potential of the secretome. The secretome, a collection of bioactive molecules secreted by cells, has shown promise in promoting tissue regeneration and modulating immune responses. This study investigates how secretome therapy can restore the integrity of keratinocytes, the primary cells responsible for maintaining the skin barrier, which is severely compromised in AD. Using in vitro AD models, the secretome's potential to reduce inflammation and enhance skin barrier function is evaluated. By targeting the underlying mechanisms of AD, secretome-based therapies could offer a novel approach to treatment, providing both regenerative and anti-inflammatory benefits. The findings from this study may pave the way for more effective, non-invasive treatments that address the root causes of AD, potentially reducing the disease's impact and improving patient outcomes.