Stem Cell Research & Therapy最新文献

{"title":"Nano-zinc oxide (nZnO) targets the AMPK-ULK1 pathway to promote bone regeneration.","authors":"Xiu Chen, Zhenkun Weng, Hongchao Zhang, Jian Jiao, Jingjia Liang, Jin Xu, Dongmei Wang, Qian Liu, Qing Yan, Aihua Gu","doi":"10.1186/s13287-025-04322-5","DOIUrl":"https://doi.org/10.1186/s13287-025-04322-5","url":null,"abstract":"<p><strong>Background: </strong>Nano-zinc oxide (nZnO) has attracted significant attention in bone tissue engineering due to its antibacterial properties, anti-inflammatory effects, biocompatibility, and chemical stability. Although numerous studies have demonstrated the enhancement of osteogenic differentiation by nZnO-modified tissue engineering materials, the underlying mechanisms remain poorly characterized.</p><p><strong>Methods: </strong>This study aimed to identify the molecular mechanisms how nZnO promoted osteogenic differentiation and bone regeneration using transcriptome analysis, drug intervention, and shRNA knockdown techniques, etc. First, the study evaluated the in vivo effects of gelatin methacryloyl (GelMA) containing nZnO on bone regeneration using a mouse calvarial defect model. The impact of nZnO exposure on the osteogenic differentiation of mesenchymal stem cells (MSCs) was then assessed. The combined treatment of nZnO and MSCs in GelMA for bone regeneration was assessed in the mouse calvarial defect model thereafter.</p><p><strong>Results: </strong>nZnO induced osteoblastic differentiation to promote bone regeneration. nZnO activated the AMP-dependent protein kinase (AMPK)-ULK1 signals to stimulate autophagosomes formation and facilitate autophagy flow, which was the essential pathway to induce osteogenic differentiation. The combined treatment of MSCs and nZnO significantly enhanced bone regeneration in calvarial defect mice. Conversely, AMPK inhibitor Compound C (C.C) reversed the effects on autophagy flow and osteogenic potentiality induced by nZnO.</p><p><strong>Conclusions: </strong>These results highlight that nZnO can regulate bone regeneration by activating autophagy through the AMPK/ULK1 signaling pathway, which may provide a novel therapeutic strategy for addressing bone defects using nZnO.</p>","PeriodicalId":21876,"journal":{"name":"Stem Cell Research & Therapy","volume":"16 1","pages":"206"},"PeriodicalIF":7.1,"publicationDate":"2025-04-24","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"https://www.ncbi.nlm.nih.gov/pmc/articles/PMC12023698/pdf/","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"144064577","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":2,"RegionCategory":"医学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"OA","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}

{"title":"Continuous collection of human mesenchymal-stromal-cell-derived extracellular vesicles from a stirred tank reactor operated under xenogeneic-free conditions for therapeutic applications.","authors":"Cristiana Ulpiano, William Salvador, Teresa Franchi-Mendes, Min-Chang Huang, Yee-Hsien Lin, Han-Tse Lin, Carlos A V Rodrigues, Ana Fernandes-Platzgummer, Joaquim M S Cabral, Gabriel A Monteiro, Cláudia L da Silva","doi":"10.1186/s13287-025-04341-2","DOIUrl":"https://doi.org/10.1186/s13287-025-04341-2","url":null,"abstract":"<p><strong>Background: </strong>Mesenchymal-stromal-cell-derived extracellular vesicles (MSC-EVs) play a key role in the paracrine effects of MSC and have demonstrated therapeutic potential in various preclinical models. However, clinical translation is hindered by manufacturing practices relying on planar culture systems, fetal bovine serum (FBS)-supplemented media, and non-scalable, low-purity EV isolation methods that fail to meet dose and safety requirements, underscoring the need for innovative approaches. In this study, we developed a scalable platform to manufacture human MSC-EVs at clinically relevant numbers, integrating continuous collection of EV-enriched conditioned media (CM) using a stirred-tank reactor (STR) under xenogeneic-free conditions and a scalable downstream process.</p><p><strong>Methods: </strong>Wharton's jelly-derived MSC (MSC(WJ)) were expanded using microcarriers in a controlled STR using human platelet lysate (hPL)-supplemented medium. Then, a 3-day EV production stage, featuring continuous harvesting of the CM, was established using a novel serum-/xeno(geneic)-free exosome depleted-hPL supplement. For the isolation of MSC-EVs, a scalable process was implemented by pairing tangential flow filtration and anion exchange chromatography. Isolated MSC-EVs were characterised using nanoparticle tracking analysis, protein and zeta potential quantification, western blot analysis of EV protein markers, transmission electron microscopy and uptake studies of fluorescently labelled-EVs.</p><p><strong>Results: </strong>The system sustained the efficient expansion of MSC(WJ), reaching a total of (6.03 ± 0.181) x 10⁷ cells after 7 days, which corresponds to a 30.1 ± 0.740-fold expansion. Upon a 3-day continuous CM harvesting, a total of (2.13 ± 0.301) x 10¹² EVs were isolated corresponding to a particle yield factor of (1.26 ± 0.186) x 10⁴ EVs/cell/day. MSC-EVs presented high purity levels ((5.53 ± 1.55) x 10⁹ particles/µg), a homogeneous small size distribution (mean diameter of 115 ± 4.88 nm), a surface charge of -23.4 ± 6.23 mV, positive detection of tetraspanins CD9 and CD63 and syntenin-1 and displayed a typical cup-shaped morphology. MSC-EVs were readily incorporated by endothelial cells and two human breast cancer cell lines.</p><p><strong>Conclusions: </strong>Overall, the scalable and Good Manufacturing Practices (GMP)-compliant platform established herein enabled the reproducible manufacturing of MSC-EVs with high purity and generally accepted characteristics concerning size, protein markers, surface charge, morphology, and cellular internalization, validating its potential for future clinical applications.</p>","PeriodicalId":21876,"journal":{"name":"Stem Cell Research & Therapy","volume":"16 1","pages":"210"},"PeriodicalIF":7.1,"publicationDate":"2025-04-24","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"https://www.ncbi.nlm.nih.gov/pmc/articles/PMC12023423/pdf/","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"143998238","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":2,"RegionCategory":"医学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"OA","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}

{"title":"PD-L1 and ICAM1 over expression empowers immunoregulation of mesenchymal stromal cells to improve the autoimmune hepatitis treatment efficacy.","authors":"Xilong Bai, Tingting Liu, Congge Li, Caie Qiu, Xiaofan Ge, Huili Gou, Hongzhi Cai, Liting Yang, Sili Wei, Wei Yang, Tianqing Li","doi":"10.1186/s13287-025-04347-w","DOIUrl":"https://doi.org/10.1186/s13287-025-04347-w","url":null,"abstract":"<p><strong>Background: </strong>Autoimmune hepatitis (AIH) is an immune-mediated disease for which there is no effective treatment. Mesenchymal stromal cells (MSCs) have become a promising treatment, but low AIH treatment efficacy has hampered the clinical application of MSCs.</p><p><strong>Methods: </strong>By using Good Manufacturing Practices, we generated mesenchymal stromal cells with enhanced immunomodulation by over-expressing PD-L1 and ICAM1 (PI-MSCs). PI-MSCs biological characteristics were established, a tertiary cell bank created, and safety of PI-MSCs determined. Finally, the efficacy of PI-MSCs for treatment of AIH was evaluated.</p><p><strong>Results: </strong>PI-MSCs preserved MSCs identity, with a normal karyotype, stable genome, and no tumorigenicity. The long-term safe dose was up to 5.0 × 10⁷ cells/kg. PI-MSCs showed better therapeutic effect than conventional MSCs for treating AIH in a mouse model. Notably, PI-MSCs showed better homing to injured liver tissue than conventional MSCs. Furthermore, PI-MSCs treatment significantly increased Treg cells in the blood and spleen of AIH model mice compared to conventional MSCs.</p><p><strong>Conclusion: </strong>PD-L1 and ICAM1 empower MSCs immuno-regulation, these empowered MSCs are more effective treatment for AIH. These findings provide support for the translation of PI-MSCs to the clinic for treatment of AIH patients.</p>","PeriodicalId":21876,"journal":{"name":"Stem Cell Research & Therapy","volume":"16 1","pages":"209"},"PeriodicalIF":7.1,"publicationDate":"2025-04-24","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"https://www.ncbi.nlm.nih.gov/pmc/articles/PMC12023376/pdf/","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"144017628","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":2,"RegionCategory":"医学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"OA","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}

{"title":"Biomanufacturing and lipidomics analysis of extracellular vesicles secreted by human blood vessel organoids in a vertical wheel bioreactor.","authors":"Justice Ene, Chang Liu, Falak Syed, Li Sun, Danyale Berry, Pradeepraj Durairaj, Zixiang Leonardo Liu, Changchun Zeng, Sunghoon Jung, Yan Li","doi":"10.1186/s13287-025-04317-2","DOIUrl":"https://doi.org/10.1186/s13287-025-04317-2","url":null,"abstract":"<p><strong>Background: </strong>Extracellular vesicles (EVs) derived from human organoids are phospholipid bilayer-bound nanoparticles that carry therapeutic cargo. However, the low yield of EVs remains a critical bottleneck for clinical translation. Vertical-Wheel bioreactors (VWBRs), with unique design features, facilitate the scalable production of EVs secreted by human blood vessel organoids (BVOs) under controlled shear stress, using aggregate- and microcarrier-based culture systems.</p><p><strong>Methods: </strong>Human induced pluripotent stem cell-derived BVOs cultured as aggregates or on Synthemax II microcarriers within VWBRs (40 and 80 rpm) were compared to static controls. The organoids were characterized by metabolite profiling, flow cytometry, and gene expression of EV biogenesis markers. EVs were characterized by nanoparticle tracking analysis, electron microscopy, and Western blotting. Lipidomics provided insights into EV lipid composition, while functional assays assessed the impact of EVs in a D-galactose-induced senescence model.</p><p><strong>Results: </strong>VWBR cultures showed more aerobic metabolism and higher expression of EV biogenesis genes compared to the static control. EVs from different conditions were comparable in size, but the yields were significantly higher for microcarrier and dynamic cultures than static aggregates. Lipidomic profiling revealed minimal variation (< 0.36%) in total lipid content; however, distinct differences were identified in lipid chain lengths and saturation levels, affecting key pathways such as sphingolipid and neurotrophin signaling. Human BVO EVs demonstrated the abilities of reducing oxidative stress and increasing cell proliferation in vitro.</p><p><strong>Conclusions: </strong>Human BVOs differentiated in VWBRs (in particular 40 rpm) produce 2-3 fold higher yield of EVs (per mL) than static control. The bio manufactured EVs from VWBRs have exosomal characteristics and therapeutic cargo, showing functional properties in in vitro assays. This innovative approach establishes VWBRs as a scalable platform for producing functional EVs with defined lipid profiles and therapeutic potential, paving the way for future in vivo studies.</p>","PeriodicalId":21876,"journal":{"name":"Stem Cell Research & Therapy","volume":"16 1","pages":"207"},"PeriodicalIF":7.1,"publicationDate":"2025-04-24","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"https://www.ncbi.nlm.nih.gov/pmc/articles/PMC12023677/pdf/","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"144039452","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":2,"RegionCategory":"医学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"OA","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}

{"title":"SERCA-mediated endoplasmic reticulum stress facilitates hematopoietic stem cell mobilization.","authors":"Lijun Li, Danhua Xu, Xinxin Huang","doi":"10.1186/s13287-025-04345-y","DOIUrl":"https://doi.org/10.1186/s13287-025-04345-y","url":null,"abstract":"<p><strong>Background: </strong>Hematopoietic stem cell (HSC) transplantation is widely recognized as an effective treatment for various malignant diseases. Enhancing HSC mobilization can improve transplantation outcomes and ultimately increase patient survival rates. Recent studies suggest that mild endoplasmic reticulum (ER) stress promotes HSC self-renewal, anti-apoptotic, and anti-aging capabilities. This led us to investigate whether inducing mild ER stress could facilitate HSC mobilization.</p><p><strong>Methods: </strong>The phenotype changes in cells treated with ER stress inducers and Sarco/endoplasmic reticulum Ca²⁺-ATPase (SERCA) inhibitors were assessed using flow cytometry. The efficacy of these agents on HSC mobilization was evaluated in C57Bl/6 mice, with colony forming unit (CFU) assays used for quantification. Knockdown Jurkat cell lines were constructed to validate the role of SERCA in the mobilization mechanism. Molecular and protein expression levels associated with the pathway were analyzed through quantitative reverse-transcription PCR and western blotting.</p><p><strong>Results: </strong>Our findings revealed that BHQ, a SERCA inhibitor, efficiently enhanced HSC mobilization in vivo. Mechanistically, BHQ regulated the CaMKII-STAT3-CXCR4 pathway by suppressing SERCA activity. This inhibition led to a reduction in CXCR4 expression on the surface of HSCs, facilitating their migration from the bone marrow into peripheral circulation.</p><p><strong>Conclusions: </strong>Our study provides novel insights into the role of the SERCA-ER stress pathway in HSC mobilization. By targeting SERCA activity with BHQ, we observed a significant enhancement in the mobilization of HSCs, facilitated by the modulation of the CaMKII-STAT3-CXCR4 signaling pathway. This research highlights the potential of utilizing mild ER stress as a strategy to promote HSC mobilization, with significant implications for improving stem cell-based therapies, including those used in HSC transplantation.</p>","PeriodicalId":21876,"journal":{"name":"Stem Cell Research & Therapy","volume":"16 1","pages":"208"},"PeriodicalIF":7.1,"publicationDate":"2025-04-24","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"https://www.ncbi.nlm.nih.gov/pmc/articles/PMC12023629/pdf/","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"144052788","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":2,"RegionCategory":"医学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"OA","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}

{"title":"Renal tubular epithelial cell-derived Exosomal miR-330-3p plays a key role in fibroblast activation and renal fibrosis by regulating CREBBP.","authors":"Rong Dai, Meng Cheng, Chu-Yi Peng, Ze-Ping Cao, Hua Jin, Dong Wang, Yi-Ping Wang, Lei Zhang","doi":"10.1186/s13287-025-04338-x","DOIUrl":"https://doi.org/10.1186/s13287-025-04338-x","url":null,"abstract":"<p><strong>Background: </strong>Renal tubular injury and activation of peripheral fibroblasts are hallmarks of chronic kidney disease (CKD), suggesting a close connection between the two cell types. Exosomes transport miRNAs and other substances to recipient cells. The involvement of exosome-mediated intercellular communication has been suggested in various diseases, including renal fibrosis. However, the underlying mechanisms remain to be determined.</p><p><strong>Methods: </strong>Rab 27a-knockout mice were constructed to confirm the role of exosomes in mice with adenine-induced renal fibrosis. Exosome secretion from the kidneys of mice with adenine-induced renal fibrosis and UA-stimulated NRK-52E cells were investigated. Exosomes released from NRK-52E cells were harvested and incubated with NRK-49 F fibroblasts or injected intravenously into the mice via the tail vein. High-throughput miRNA sequencing was used to evaluate the miRNA profiles of UA-Exo. The roles of candidate miRNAs and their target genes and related pathways were predicted and evaluated in vitro and in vivo using specific miRNA mimics, miRNA inhibitors, siRNAs, and adeno-associated viruses (AAV).</p><p><strong>Results: </strong>Inhibition of exosome secretion by Rab27a knockout or siRNA Rab27a treatment inhibited fibroblast activation and ameliorated renal fibrosis. Significantly increased renal fibrosis was seen in mice treated with adenine, and exosome release was increased after UA treatment of NRK-52E cells. Exosomes released from NRK-52E cells after UA stimulation activated fibroblasts and exacerbated renal fibrosis. The expression of miR-330-3p in exosomes was significantly increased in the UA-Exo group compared with the control group, suggesting the potential use of miR-330-3p as a marker of renal fibrosis. CREBBP was found to be a specific target of miR-330-3p. The stimulation or inhibition of exosomal miR-330-3p release from renal tubular epithelial cells thus promoted or blocked fibroblast activation in vitro, while miR-330-3p-deficient exosomes attenuated renal fibrosis by modulating CREBBP in vivo.</p><p><strong>Conclusion: </strong>The findings suggest that exosomes play an important role in promoting renal fibrosis by mediating the communication between renal tubular epithelial cells and fibroblasts. This role is associated with inhibition of CREBBP activity by exosomal miR-330-3p in fibroblasts. Thus, the miR-330-3p/CREBBP axis is a promising target for the treatment and management of renal fibrosis.</p>","PeriodicalId":21876,"journal":{"name":"Stem Cell Research & Therapy","volume":"16 1","pages":"203"},"PeriodicalIF":7.1,"publicationDate":"2025-04-23","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"https://www.ncbi.nlm.nih.gov/pmc/articles/PMC12020049/pdf/","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"144039121","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":2,"RegionCategory":"医学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"OA","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}

{"title":"A first-in-human, prospective pilot trial of umbilical cord-derived mesenchymal stem cell eye drops therapy for patients with refractory non-Sjögren's and Sjögren's syndrome dry eye disease.","authors":"Di Zhang, Taige Chen, Qi Liang, Xuebing Feng, Jiaxuan Jiang, Zeying Chen, Yun Tang, Yiran Chu, Bin Wang, Kai Hu","doi":"10.1186/s13287-025-04292-8","DOIUrl":"10.1186/s13287-025-04292-8","url":null,"abstract":"&lt;p&gt;&lt;strong&gt;Background: &lt;/strong&gt;Patients with refractory dry eye disease (DED) often face the threat of diminished visual quality and have limited responses to existing treatments. Ocular injection of Mesenchymal stem cells (MSCs) has recently emerged as a promising new therapeutic strategy for DED. Topical eye drops are the clinical favorable choice for drug administration in DED. To date, the clinical use of MSC eye drops has not been reported in settings. This clinical trial represents a groundbreaking exploration into the preliminary therapeutic potential and safety of umbilical cord MSC eye drops for patients with refractory DED, including both non-Sjögren's dry eye (NSDE) and Sjögren's syndrome dry eye (SSDE). The study also aimed to investigate the possible underlying mechanisms.&lt;/p&gt;&lt;p&gt;&lt;strong&gt;Methods: &lt;/strong&gt;In this open-label, prospective, single-arm, self-controlled trial, 11 NSDE and 5 SSDE patients received twice-daily MSC eye drops for two weeks, subsequent follow-up visits were scheduled at 4 weeks and 12 months after treatment. The primary efficacy was evaluated using the ocular surface disease index (OSDI) score, tear meniscus height (TMH), non-invasive break-up time (NIBUT), Schirmer I test (SIT), and corneal fluorescein staining (CFS) score. Secondary assessments focused on the evaluation of lipid layer, meibomian gland function, and bulbar conjunctival redness. Safety was monitored by recording adverse events (AEs) throughout the study. Changes in tear levels of interleukin-6 (IL-6), IL-17A, Mucin 5AC (MUC5AC), C-C chemokine ligand 20 (CCL20) and IL-23, along with proteomic alterations, were compared between baseline and T-week2.&lt;/p&gt;&lt;p&gt;&lt;strong&gt;Results: &lt;/strong&gt;Significant clinical improvements were observed in most symptoms and signs following MSC eye drops treatment in both NSDE and SSDE patients, particularly in tear production as measured by SIT and TMH, and the alleviation of meibomian gland blockage. The therapeutic effect on OSDI, NIBUT, and the lipid layer was more pronounced in NSDE patients compared to SSDE. No serious AEs were reported during the treatment and follow-up period. Post-treatment reductions in tear levels of IL-6 and IL-17A, along with an increase in MUC5AC, further confirmed the efficacy. Tear proteomic analysis indicated that the efficacy of MSC eye drops is associated with the inhibition of inflammation caused by T helper 17 (Th17) cells in both NSDE and SSDE groups.&lt;/p&gt;&lt;p&gt;&lt;strong&gt;Conclusions: &lt;/strong&gt;In this prospective exploratory clinical study, we have demonstrated that MSC eye drops might offer clinical efficacy and manageable safety in treating refractory DED for the first time, potentially bringing a new perspective on the treatment of such patients. Our research represents a preliminary exploratory endeavor, paving the way for future large-scale randomized positive-controlled trials.&lt;/p&gt;&lt;p&gt;&lt;strong&gt;Trial registration: &lt;/strong&gt;ClinicalTrials.gov, NCT05784519. Registered 28 February 2023, htt","PeriodicalId":21876,"journal":{"name":"Stem Cell Research & Therapy","volume":"16 1","pages":"202"},"PeriodicalIF":7.1,"publicationDate":"2025-04-23","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"https://www.ncbi.nlm.nih.gov/pmc/articles/PMC12020048/pdf/","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"144064498","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":2,"RegionCategory":"医学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"OA","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}

{"title":"Sildenafil promotes osteogenic differentiation of human mesenchymal stem cells and inhibits bone loss by affecting the TGF-β signaling pathway.","authors":"Menglong Hu, Likun Wu, Erfan Wei, Xingtong Pan, Qiyue Zhu, Xv Xiuyun, Letian Lv, Xinyi Dong, Hao Liu, Yunsong Liu","doi":"10.1186/s13287-025-04320-7","DOIUrl":"https://doi.org/10.1186/s13287-025-04320-7","url":null,"abstract":"<p><strong>Background: </strong>Osteoporosis, a common bone disorder, is primarily managed pharmacologically. However, existing medications are associated with non-trivial side-effects. Sildenafil, which already finds many clinical applications, promotes angiogenesis and cellular differentiation. Osteoporotic patients often exhibit a reduced intraosseous vasculature and impaired cellular differentiation; sildenafil may thus usefully treat osteoporosis.</p><p><strong>Methods: </strong>Here, the effects of sildenafil on the osteogenic differentiation of human mesenchymal stem cells (hMSCs) were explored, as were the molecular mechanisms in play. We treated hMSCs with varying concentrations of sildenafil and measured cell proliferation and osteogenic differentiation in vitro. We used a mouse model of subcutaneous ectopic osteogenesis to assess sildenafil's effect on hMSC osteogenic differentiation in vivo. We also explored the effects of sildenafil on bone loss in tail-suspended (TS) and ovariectomized (OVX) mice. Mechanistically, we employed RNA-sequencing to define potentially relevant molecular pathways.</p><p><strong>Results: </strong>The appropriate concentrations of sildenafil significantly enhanced osteogenic hMSC differentiation; the optimal sildenafil concentration may be 10 mg/L. Sildenafil mitigated osteoporosis in OVX and TS mice. The appropriate concentrations of sildenafil probably promoted hMSC osteogenic differentiation by acting on the transforming growth factor-β (TGF-β) signaling pathway.</p><p><strong>Conclusions: </strong>In conclusion, sildenafil enhanced hMSC osteogenic differentiation and inhibited bone loss. Sildenafil may usefully treat osteoporosis. Our findings offer new insights into the physiological effects of the medicine.</p>","PeriodicalId":21876,"journal":{"name":"Stem Cell Research & Therapy","volume":"16 1","pages":"201"},"PeriodicalIF":7.1,"publicationDate":"2025-04-23","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"https://www.ncbi.nlm.nih.gov/pmc/articles/PMC12016470/pdf/","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"143997732","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":2,"RegionCategory":"医学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"OA","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}

{"title":"Human platelet lysate produced from leukoreduction filter contents enables sufficient MSC growth.","authors":"Shinobu Wakamoto, Tomoko Furukawa, Masahito Kawabori, Mitsuaki Akino, Shiho Kato, Hisae Fuse, Sumio Ohtsuki, Yoshihiro Torimoto, Miki Fujimura, Shuichi Kino","doi":"10.1186/s13287-025-04329-y","DOIUrl":"https://doi.org/10.1186/s13287-025-04329-y","url":null,"abstract":"&lt;p&gt;&lt;strong&gt;Background: &lt;/strong&gt;Stem cell therapy holds significant potential for promoting recovery, with numerous products currently under development. Blood-derived supplements are often essential for successful stem cell expansion, with fetal bovine serum (FBS) being the most commonly used supplement. However, FBS has drawbacks, including the risk of immune responses, ethical concerns about animal welfare, and potential zoonotic infections. Human platelet lysate (hPL), derived from lysed platelets, contains various growth factors and has been proposed as an alternative to FBS. However, obtaining sufficient human platelets for clinical use remains challenging. Leukoreduction filters, used during blood transfusion manufacturing to remove leukocytes, also retain significant amounts of platelets and plasma. This study investigates the feasibility and efficacy of filter-derived hPL (f-hPL) for mesenchymal stem cell (MSC) expansion.&lt;/p&gt;&lt;p&gt;&lt;strong&gt;Methods: &lt;/strong&gt;Leukoreduction filters were collected after their use in the manufacturing of whole blood transfusion products. Each filter was reverse-perfused with saline to extract residual blood contents. Platelets (f-platelet) and supernatant were separated by multiple centrifugation steps. f-Platelet were lysed with varying concentrations of fresh frozen plasma (FFP) to determine the optimal protein concentration for the lysate solution. Then, plasma left in the leukoreduction filters were used to generate lysate solution (f-plasma) at optimal protein concentration. f-Platelet (1.1 × 10&lt;sup&gt;9&lt;/sup&gt;/mL) and f-plasma (27 mg/mL protein) were combined in a freezing bag and subjected to three freeze-thaw cycles to produce f-hPL. Both small- and large-scale f-hPL were manufactured, and MSCs expansion and quality assessments were perfomed to evaluate the efficacy of f-hPL.&lt;/p&gt;&lt;p&gt;&lt;strong&gt;Results: &lt;/strong&gt;A total of 3.5 ± 0.6 × 10&lt;sup&gt;10&lt;/sup&gt; f-platelets were obtained from a single leukoreduction filter, yielding a collection efficiency of 37.1 ± 5.3%. The optimal protein concentration of lysate solution for cell expansion was &gt; 27 mg/mL. Subsequently, six leukoreduction filters used to produce enough f-platelet and p-plasma for 100 mL of f-hPL. MSCs cultured in medium supplemented with 10% f-hPL demonstrated superior expansion, with cell proliferation rates 20% higher than those observed with commercial hPL and 300% higher than those cultured with FBS. The expanded MSCs met the International Society for Cell & Gene Therapy criteria for cell surface markers and differentiation potential. MSCs expanded with f-hPL expressed similar to or higher amounts of hepatocyte growth factor compared with those cultured with FBS and human AB serum. Furthermore, f-hPL significantly enhanced cell proliferation up to P12 and effectively prevented cell senescence.&lt;/p&gt;&lt;p&gt;&lt;strong&gt;Conclusion: &lt;/strong&gt;f-hPL derived from leukoreduction filters demonstrated strong capacity for MSCs expansion. The use of discarded blood material","PeriodicalId":21876,"journal":{"name":"Stem Cell Research & Therapy","volume":"16 1","pages":"205"},"PeriodicalIF":7.1,"publicationDate":"2025-04-23","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"https://www.ncbi.nlm.nih.gov/pmc/articles/PMC12020118/pdf/","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"144000112","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":2,"RegionCategory":"医学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"OA","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}

{"title":"LncRNA MRF targeting FSHR inhibits the osteogenic differentiation of BMSCs and bone defect repair through the regulation of the cAMP-PKA-CREB signaling pathway.","authors":"Qing Ning, Ming Li, Zhuangyao Liao, Enming Chen, Huatao Liu, Yuwei Liang, Yuanquan Chen, Yuxi Li, Lin Huang","doi":"10.1186/s13287-025-04291-9","DOIUrl":"https://doi.org/10.1186/s13287-025-04291-9","url":null,"abstract":"<p><strong>Background: </strong>Mesenchymal stem cells (MSCs), known for their ability to differentiate into osteoblasts, play a pivotal role in bone metabolism. In our previous investigations, we identified a novel long non-coding RNA (lncRNA) named MCP1 Regulatory Factor (MRF), which exhibits significant involvement in immune regulation of BMSCs. Moreover, we observed noticeable expression changes of MRF during the osteogenic differentiation of BMSCs. However, the exact role and underlying mechanism of MRF in the osteogenic differentiation of BMSCs remain elusive.</p><p><strong>Methods: </strong>QRT-PCR analysis was employed to assess the expression levels of MRF. RNA interference and overexpression plasmids were utilized to modulate MRF expression, allowing for the observation of changes in the osteogenic differentiation capacity of BMSCs. Downstream pathways involved in the MRF-mediated regulation of BMSCs' osteogenic differentiation were predicted using transcriptome sequencing. The functionality of MRF in vivo was validated through a mouse tibial drilling defect model.</p><p><strong>Results: </strong>In patients with osteoporosis, there is a notable increase in the expression of MRF within BMSCs. During the osteogenic differentiation of BMSCs, the MRF expression progressively decreases. The knockdown of MRF significantly enhances the osteogenic differentiation of BMSCs, promoting an increased expression of bone-related proteins such as RUNX2, ALP, and COL1A1. Transcriptome sequencing and western blot indicated that cAMP/PKA/CREB signaling pathway was significantly activated after lncRNA-MRF knockdown. Moreover, in the mouse tibial drilling defect model, MRF knockdown significantly promotes ossification in vivo.</p><p><strong>Conclusions: </strong>MRF modulates the cAMP/PKA/CREB signaling pathway via the follicle stimulating hormone receptor (FSHR), thereby influencing the ossification differentiation of BMSCs. Our research suggests that MRF may serve as a potential target for bone-related disorders.</p>","PeriodicalId":21876,"journal":{"name":"Stem Cell Research & Therapy","volume":"16 1","pages":"200"},"PeriodicalIF":7.1,"publicationDate":"2025-04-23","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"https://www.ncbi.nlm.nih.gov/pmc/articles/PMC12016372/pdf/","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"144047543","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":2,"RegionCategory":"医学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"OA","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}