Regenerative Therapy最新文献

{"title":"Dose-response immunomodulatory effects of Mesenchymal stem cell-derived culture-conditioned media in acute Graft-versus-Host Disease","authors":"Mohini Mendiratta ,&nbsp;Meenakshi Mendiratta ,&nbsp;Sujata Mohanty ,&nbsp;Sandeep Rai ,&nbsp;Ritu Gupta ,&nbsp;Vatsla Dadhwal ,&nbsp;Sameer Bakhshi ,&nbsp;Deepam Pushpam ,&nbsp;Mukul Aggarwal ,&nbsp;Aditya Kumar Gupta ,&nbsp;Ranjit Kumar Sahoo","doi":"10.1016/j.reth.2025.10.001","DOIUrl":"10.1016/j.reth.2025.10.001","url":null,"abstract":"<div><h3>Background</h3><div>Mesenchymal stem cell-based therapy faces challenges that have driven interest in MSCs-derived culture-conditioned media (CCM) as a cell-free alternative. Our study aims to optimize the dose and collection timing of CCM to enhance its therapeutic efficacy in aGVHD, while also standardizing co-culture conditions for CD3⁺ T-cell interaction with CCM.</div></div><div><h3>Material and methods</h3><div>Human MSCs were isolated from BM and WJ and subsequently preconditioned under hypoxic conditions (1 % O₂) for 24 h in a tri-gas incubator. Culture-conditioned media (CCM) were collected from both naive (MSCs) and hypoxia-preconditioned MSCs (MSCs^HYP) at 24, 48, and 72 h and filtered using a 0.2 μm membrane filter. CD3⁺ T-cell were isolated from PBMNCs derived from aGVHD patients. These T-cell were co-cultured at varying densities (2∗10⁶, 5∗10⁶, and 10∗10⁶ cells/ml) with different concentrations of CCM (25 %, 50 %, and 100 %), and cell proliferation was assessed using the MTS assay. Furthermore, CD3⁺ T-cell proliferation and activation status were evaluated in a 2D co-culture model of CD3⁺ T-cell and CCM using flow cytometry.</div></div><div><h3>Results</h3><div>Our findings revealed that CCM collected at 48 h, at a 50 % concentration, exerted the most pronounced inhibitory effect on CD3⁺ T-cell proliferation, particularly at a density of 5∗10⁶ cells/ml, irrespective of the MSCs source. Hypoxia preconditioning significantly enhanced the immunomodulatory effects, with WJ-MSCs^HYP-CCM demonstrating superior efficacy in suppressing T-cell proliferation, increasing the CD4⁺/CD8⁺ T-cell ratio, and reducing CD4⁺ T-cell activation compared to BM-MSCs^HYP-CCM.</div></div><div><h3>Conclusion</h3><div>These results emphasize the critical role of optimizing CCM collection timing and concentration to maximize therapeutic potential. Our study paves the way for the development of standardized, scalable, and effective cell-free therapies for aGVHD.</div></div>","PeriodicalId":20895,"journal":{"name":"Regenerative Therapy","volume":"30 ","pages":"Pages 868-874"},"PeriodicalIF":3.5,"publicationDate":"2025-10-08","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"145265262","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":3,"RegionCategory":"环境科学与生态学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}

{"title":"Modeling epithelial wound closure dynamics with AI: A comparative study across cell types","authors":"Xueyao Cai ,&nbsp;Weidong Li ,&nbsp;Wenjun Shi ,&nbsp;Yuchen Cai ,&nbsp;Jianda Zhou","doi":"10.1016/j.reth.2025.10.002","DOIUrl":"10.1016/j.reth.2025.10.002","url":null,"abstract":"<div><h3>Background</h3><div>Skin wound healing exhibits complex spatiotemporal heterogeneity that challenges traditional static assessment methods. Current artificial intelligence (AI) approaches often treat segmentation and temporal modeling as disconnected processes, limiting dynamic quantification of healing trajectories across distinct cell types. We aim to develop an integrated AI framework combining enhanced segmentation with temporal modeling for quantifying <em>in vitro</em> wound closure dynamics in normal epithelial (MCF10A) and tumor (MCF7) cells, and to compare algorithmic performance across healing phenotypes.</div></div><div><h3>Methods</h3><div>We implemented an enhanced UNet++ model for wound segmentation in time-lapse images, benchmarked against Otsu thresholding. Temporal closure trajectories were modeled using polynomial regression, Random Forest (RF), Support Vector Regression (SVR), Autoregressive Integrated Moving Average (ARIMA), and Temporal Convolutional Network (TCN). Performance was evaluated via Dice/IoU (segmentation) and MAE/R² (temporal modeling).</div></div><div><h3>Results</h3><div>UNet++ achieved significantly higher segmentation accuracy than Otsu thresholding (Dice: <em>p</em> = 8.841 × 10⁻⁴⁹; IoU: <em>p</em> = 3.931 × 10⁻⁴⁷) with consistent temporal robustness across healing phases. For closure trajectory modeling, RF achieved superior accuracy for MCF7 (mean absolute error [MAE] = 0.48 %, R² = 0.968) and MCF10A (MAE = 1.73 %, R² = 0.872), excelling in capturing nonlinear phase transitions and plateau behaviors. TCN showed promise for abrupt changes in MCF7 (MAE = 1.67 %, R² = 0.698) but failed for near-stationary MCF10A trends. Significant cell-type differences emerged, with RF providing the most interpretable predictions.</div></div><div><h3>Conclusion</h3><div>This integrated framework enables precise dynamic wound monitoring, holding clinical potential for chronic ulcer management and tumor margin surveillance, particularly through its ability to discern cell-type-specific healing phenotypes.</div></div>","PeriodicalId":20895,"journal":{"name":"Regenerative Therapy","volume":"30 ","pages":"Pages 860-867"},"PeriodicalIF":3.5,"publicationDate":"2025-10-04","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"145219334","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":3,"RegionCategory":"环境科学与生态学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}

{"title":"No simple way to averaging out: Pooled mesenchymal stromal cells do not reflect average donor characteristics","authors":"Dea Kukaj ,&nbsp;Sabine Niebert ,&nbsp;Christoph Biehl ,&nbsp;Ursula Reichart ,&nbsp;Christiane Schueler ,&nbsp;Janina Burk","doi":"10.1016/j.reth.2025.09.012","DOIUrl":"10.1016/j.reth.2025.09.012","url":null,"abstract":"<div><h3>Background</h3><div>Mesenchymal stromal cells (MSCs) are promising candidates for numerous regenerative therapies. Still, clinical translation is complicated by the heterogeneity of MSCs and related shortcomings in preclinical research. Pooling MSCs from multiple donors is increasingly being advocated as an effective way to mitigate donor variability. However, it remains unclear whether the range of individual cell characteristics is equally reflected in pooled cultures, or if pooling rather leads to a homogenized cell population dominated by the fittest donor, which would lead to skewed results. This study investigates whether MSC pools are functionally representative for their respective donor MSCs and whether dominant donors emerge over time.</div></div><div><h3>Methods</h3><div>MSCs from nine human donors were categorized into low-, middle-, and high-fitness groups. Individual MSCs were then pooled according to their fitness groups, complemented by a mixed-fitness pool. Functional assays for proliferation, metabolic activity, differentiation, migration and senescence were performed to evaluate the pools versus the individual MSCs. Donor representation within pools was tracked using fluorescence microscopy and qPCR.</div></div><div><h3>Results</h3><div>The high-fitness pool, as well as its individual donor MSCs, displayed the most rapid proliferation and highest metabolic activity. However, while for proliferation, the pool data aligned well with the individual donor data, all other assays revealed discrepancies between the pooled cultures and individual donor cells. Interestingly, particularly the mixed fitness pool showed inferior metabolic activity and differentiation potential in comparison with the respective individual donor MSCs. Cell tracking showed that over one passage, even pools composed of donors with similar cell fitness became dominated by the donor with the highest cellular fitness.</div></div><div><h3>Conclusions</h3><div>The discrepancy between pooled and individual donor data emphasizes the importance of biological replicates to capture donor variation and ensure that MSC research reflects natural diversity.</div></div>","PeriodicalId":20895,"journal":{"name":"Regenerative Therapy","volume":"30 ","pages":"Pages 849-859"},"PeriodicalIF":3.5,"publicationDate":"2025-10-03","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"145219332","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":3,"RegionCategory":"环境科学与生态学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}

{"title":"NN15-017 promotes human pluripotent stem cell proliferation and generation","authors":"Shizuka Fujigaki-Yamazaki ,&nbsp;Ryoko Nakatani ,&nbsp;Osamu Hirata ,&nbsp;Koichiro Saruhashi ,&nbsp;Mai Fujii ,&nbsp;Hirofumi Suemori ,&nbsp;Taito Nishino ,&nbsp;Eihachiro Kawase","doi":"10.1016/j.reth.2025.09.003","DOIUrl":"10.1016/j.reth.2025.09.003","url":null,"abstract":"<div><h3>Introduction</h3><div>The nutrient requirements for culturing human pluripotent stem cells (hPSCs) have been studied in relation to the development of chemically defined media. Most hPSC media contain FGF2, which is critical for hPSC growth and maintenance. Here, we investigated compounds that could substitute for FGF2 activity.</div></div><div><h3>Methods</h3><div>To identify compounds that can replace FGF2 and promote the proliferation of undifferentiated hPSCs, we screened our compound library using the hOCT4-EGFP reporter system and evaluated candidates using cell morphological analysis and OCT4 immunostaining as a marker of undifferentiated cells. We further evaluated the extent to which the selected compound can replace FGF2 through long-term culture (over 10 passages). We examined whether the compounds promote the reprogramming efficiency of somatic cells into induced pluripotent stem cells (iPSCs) using mRNA reprogramming. We also analyzed which signaling pathways were activated by the compound using western blotting.</div></div><div><h3>Results</h3><div>We identified a new compound, NN15-017, which enables a reduced concentration of FGF2 in the medium by 5-fold and enhances the reprogramming efficiency of human induced PSCs by 2- to 3-fold. NN15-017 promoted the MAP/ERK signaling pathway downstream of FGF2 and may affect the Hippo-YAP signaling pathway in hPSCs.</div></div><div><h3>Conclusions</h3><div>NN15-017 reduces the requirement for FGF2, thereby providing novel and valuable benefits for the proliferation and generation of human induced PSCs.</div></div>","PeriodicalId":20895,"journal":{"name":"Regenerative Therapy","volume":"30 ","pages":"Pages 838-848"},"PeriodicalIF":3.5,"publicationDate":"2025-10-02","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"145219349","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":3,"RegionCategory":"环境科学与生态学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}

{"title":"A comprehensive review of clinical trials and Progress in stem cell therapies for advanced heart failure","authors":"Thi-Thu-Hien Tran ,&nbsp;Thuy-Linh Tran ,&nbsp;Ha-Tung-Chi Nguyen ,&nbsp;Thi-Hoa Luong ,&nbsp;Thi-Minh-Anh Dang ,&nbsp;Tuan-Anh Pham ,&nbsp;Thi-Nhung Nguyen","doi":"10.1016/j.reth.2025.09.009","DOIUrl":"10.1016/j.reth.2025.09.009","url":null,"abstract":"<div><div>Heart failure (HF) is a global epidemic with a rising burden on individuals and healthcare systems. Advanced HF is characterized by severe symptoms at rest and marked limitations in physical activity. Stem cell therapies potentially address the limitations of current conventional treatments by reversing damage to cardiac tissue via their unique capacity for self-renewal and multilineage differentiation.</div><div>This article summarized the mechanism of action and methods for each stem cell approach and provided a detailed summary of associated clinical trials focusing on efficacy, safety profiles, and future directions.</div><div>We conducted a systematic review with a comprehensive search from 2014 to 2024 using databases such as PubMed, SCOPUS, and Google Scholar. Articles that met the inclusion criteria were selected by abstract review and subsequent assessment of the full text.</div><div>This review analyzes 27 clinical trials investigating stem cell-based therapies for advanced HF across different clinical phases (Phases I, II, and III) conducted between 2014 and 2024. These approaches include adult stem cells (ASCs) such as cardiac stem cells (CSCs), cardiosphere-derived cells (CDCs), cardiac progenitor cells (CPCs), unfractionated bone marrow-derived mononuclear cells (BMMNCs), mesenchymal stem cells (MSCs) and pluripotent stem cells (PSCs) such as embryonic stem cells (ESCs, induced pluripotent stem cells (iPSCs).</div><div>While the exact mechanisms through which stem cells exert their therapeutic effects in advanced HF remain under investigation, recent attention has shifted toward the paracrine signaling effects of injected cells. All approaches have demonstrated clinically acceptable safety profiles; however, their efficacy varies and has yet to be conclusively confirmed. MSC-based therapy is the most widely used among the different cell choices and has consistently exhibited promising outcomes. Although ESCs are promising for heart regeneration, their use is significantly limited by ethical issues that advancements in iPSCs may potentially address. With further efficacy validation, especially in phase III clinical trials, stem cell therapies hold promising potential for clinical application in advanced HF.</div></div>","PeriodicalId":20895,"journal":{"name":"Regenerative Therapy","volume":"30 ","pages":"Pages 812-837"},"PeriodicalIF":3.5,"publicationDate":"2025-10-01","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"145219331","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":3,"RegionCategory":"环境科学与生态学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}

{"title":"Research trends on platelet-rich plasma in improving ovarian dysfunction: A bibliometric and visualization analysis","authors":"Fangyu Liu ,&nbsp;Jiajia Li ,&nbsp;Jin Li ,&nbsp;Maozhuo Lan ,&nbsp;Xiaoning Wang ,&nbsp;Songling Zhang","doi":"10.1016/j.reth.2025.09.010","DOIUrl":"10.1016/j.reth.2025.09.010","url":null,"abstract":"<div><h3>Introduction</h3><div>Platelet-rich plasma (PRP) has attracted attention due to its potential to restore or enhance ovarian function. However, resources for evaluating the research trends of PRP in improving ovarian dysfunction are limited. This study aims to conduct a bibliometric analysis of the relevant literature on PRP improving ovarian dysfunction, and explore the current research status, hotspots and frontiers in this field.</div></div><div><h3>Methods</h3><div>Studies on PRP improving ovarian dysfunction were retrieved from the Science Citation Index Extended (SCI-E) database of Web of Science (WOS). The research trends and characteristics were analyzed using visualization software such as VOSviewer, SCImago Graphica and CiteSpace.</div></div><div><h3>Results</h3><div>A total of 193 studies were identified. The USA, IRAN and CHINA were the most productive countries. The number of publications on PRP improving ovarian dysfunction has shown a significant upward trend since 2018. Papers published in the United States lead in terms of the number of publications, citation counts and H-index. Although the number of publications in Turkey is slightly lower than that in China, its H-index and average citations are higher than those in China. The strongest keyword is “stimulation”, and the keyword with the longest duration of an outbreak is “gonadotropins” (starting in 2010 and ending in 2018). The 5 largest co-citation clusters are as follows: menopause (#0), fertility preservation (#1), implantation failure (#3), preimplantation genetic testing (#4), stem cell therapy (#5).</div></div><div><h3>Conclusions</h3><div>Current research indicates that there is still great potential for the development of PRP in improving ovarian dysfunction. In the future, it is necessary to break through the current bottleneck, promote the transformation of PRP from a “potential intervention method” to a “clinical routine plan”, and provide safer and more effective treatment options for patients with decreased ovarian dysfunction and assisted reproduction.</div></div>","PeriodicalId":20895,"journal":{"name":"Regenerative Therapy","volume":"30 ","pages":"Pages 802-811"},"PeriodicalIF":3.5,"publicationDate":"2025-09-30","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"145219330","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":3,"RegionCategory":"环境科学与生态学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}

{"title":"Advancing the understanding of alveolar regeneration: Global research trends, thematic evolution, and emerging frontiers","authors":"Si-Cheng Zhou ,&nbsp;Ying-Xian Dong ,&nbsp;Jie Tian ,&nbsp;Guo-Wei Che ,&nbsp;Yutian Lai","doi":"10.1016/j.reth.2025.09.005","DOIUrl":"10.1016/j.reth.2025.09.005","url":null,"abstract":"<div><h3>Background</h3><div>Alveolar regeneration represents a critical research direction in respiratory disease treatment. Despite the surge in studies following the COVID-19 pandemic, comprehensive bibliometric analysis to systematically evaluate global research trends and future directions remains lacking<strong>.</strong></div></div><div><h3>Methods</h3><div>This study employed bibliometric methodology to analyze 1564 publications related to alveolar regeneration from 1974 to 2024 using the Web of Science Core Collection database. Data visualization and analysis were conducted using VOSviewer (version 1.6.19), CiteSpace (version 6.2.R3), and the biblioshiny R package.</div></div><div><h3>Results</h3><div>The analysis encompassed 68 countries, 1930 institutions, and 9150 researchers across 658 journals. The United States leads with 601 publications and 32,172 citations, with Harvard University as the most influential institution. The <em>American Journal of Respiratory and Critical Care Medicine</em> has the highest impact factor (19.3), while the <em>American Journal of Physiology-Lung Cellular and Molecular Physiology</em> has the most co-citations (2,402). Edward E. Morrisey is the most prolific author, and C. E. Barkauskas has the highest co-citations. Keyword analysis revealed six major research clusters: stem cells and regenerative medicine, acute lung injury and fibrosis, COVID-19-related research, chronic lung disease repair, cellular behavior and molecular mechanisms, and post-pneumonectomy regeneration. Thematic mapping indicates future research should prioritize lung injury repair mechanisms, matrix environment in tissue regeneration, stem cell therapeutics, and immune regulation in lung injury repair.</div></div><div><h3>Conclusion</h3><div>This first comprehensive 50-year bibliometric analysis of alveolar regeneration reveals the evolutionary trend from basic mechanistic exploration toward clinical translational applications, providing important reference for researchers and funding agencies.</div></div>","PeriodicalId":20895,"journal":{"name":"Regenerative Therapy","volume":"30 ","pages":"Pages 778-794"},"PeriodicalIF":3.5,"publicationDate":"2025-09-30","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"145219333","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":3,"RegionCategory":"环境科学与生态学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}

{"title":"Anastomotic leakage prevention using dry-preserved fibroblast cell sheets in esophageal reconstruction","authors":"Hiroshi Kurazumi ,&nbsp;Ryunosuke Sakamoto ,&nbsp;Koji Ueno ,&nbsp;Akihiro Fujita ,&nbsp;Kazumasa Matsunaga ,&nbsp;Masashi Yanagihara ,&nbsp;Yoshihiro Takemoto ,&nbsp;Junichi Murakami ,&nbsp;Atsunori Oga ,&nbsp;Shunsaku Katsura ,&nbsp;Kimikazu Hamano","doi":"10.1016/j.reth.2025.09.011","DOIUrl":"10.1016/j.reth.2025.09.011","url":null,"abstract":"<div><h3>Background</h3><div>Anastomotic leakage is a common and serious complication of esophageal reconstruction, and new methods are required for its prevention in clinical settings. We herein developed dry-preserved fibroblast cell sheets (dry sheets), which are easy to use and promote wound healing. This study aimed to investigate the efficacy of transplanting allogeneic dry sheets in preventing anastomotic leakage in a rat esophageal reconstruction model.</div></div><div><h3>Methods</h3><div>Allogeneic dry sheets were prepared from the rat oral mucosa. A rat esophageal anastomosis model was created, and two dry sheets were applied to cover the anastomotic sites. Anastomotic leakage incidence, burst pressure, histological findings, and collagen contents were compared between the control and dry sheet groups postoperatively.</div></div><div><h3>Results</h3><div>The dry sheet group demonstrated a lower incidence of anastomotic leakage than the control group (control: 64 % vs. dry sheet: 28 % on day 3, control: 57 % vs. dry sheet: 29 % on day 5). Abscess scores at the esophageal anastomotic sites were also lower in the dry sheet group than in the control group on days 3 and 5. Burst pressure was significantly higher in the dry sheet group than in the control group on days 3 and 5. Collagen type I was significantly increased in the dry sheet group compared with that in the control group.</div></div><div><h3>Conclusions</h3><div>Allogeneic dry sheet application improved anastomotic leakage incidence and burst pressure, indicating the usefulness of these sheets in preventing esophageal anastomotic leakage.</div></div>","PeriodicalId":20895,"journal":{"name":"Regenerative Therapy","volume":"30 ","pages":"Pages 795-801"},"PeriodicalIF":3.5,"publicationDate":"2025-09-30","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"145219329","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":3,"RegionCategory":"环境科学与生态学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}

{"title":"Cholangiocyte-derived exosomal miR-381-3p promotes hepatic stellate cell activation and cholestatic liver fibrosis via targeting Klf6","authors":"Yang Gao,&nbsp;Yonglin Chen,&nbsp;Yuanyi Mang,&nbsp;Xibing Zhang,&nbsp;Xiaoshan Li,&nbsp;Shengning Zhang","doi":"10.1016/j.reth.2025.09.008","DOIUrl":"10.1016/j.reth.2025.09.008","url":null,"abstract":"<div><h3>Introduction</h3><div>The activation of hepatic stellate cells (HSCs) is a central mechanism driving the progression of chronic cholestatic liver diseases (CCLD). MicroRNAs (miRNAs) have been increasingly recognized for their regulatory roles in various liver pathologies. This study aimed to identify specific miRNAs involved in the progression of CCLD and elucidate their underlying molecular mechanisms.</div></div><div><h3>Methods</h3><div>A bile duct ligation (BDL) mouse model was established to mimic cholestatic liver injury. Exosomes were isolated from mouse large cholangiocytes (MLE) and co-cultured with HSCs to investigate their functional effects. Reverse transcription quantitative polymerase chain reaction (RT-qPCR) was employed to assess the expression levels of fibrotic marker genes. Exosomal miRNA microarray assay was used to screen differentially expressed miRNAs. Furthermore, dual-luciferase reporter assays and RNA pull-down experiments were conducted to validate the interaction between miR-381-3p and its putative target gene, kinesin family member 6 (Klf6).</div></div><div><h3>Results</h3><div>The results confirmed the successful isolation of exosomes from MLE. Furthermore, exosomes derived from BDL-MLE were shown to promote the activation of HSCs and exacerbate hepatic fibrosis. Dicer knockout reduced the mRNA expression of key fibrotic markers. In addition, miR-381-3p was found to be upregulated both in BDL-MLE–derived exosomes and in HSCs treated with transforming growth factor (TGF)-β and BDL-MLE–exosomes. Mechanistic investigations identified Klf6 as a direct target gene of miR-381-3p. Rescue experiments further revealed that overexpression of Klf6 alleviated the pro-fibrotic effects caused by miR-381-3p overexpression, thereby attenuating HSC activation and mitigating cholestatic liver fibrosis.</div></div><div><h3>Conclusion</h3><div>This study highlighted the role of MLE-derived exosomal miR-381-3p in promoting HSC activation and cholestatic liver fibrosis through the regulation of Klf6. These findings provide novel insights into the molecular mechanisms underlying CCLD.</div></div>","PeriodicalId":20895,"journal":{"name":"Regenerative Therapy","volume":"30 ","pages":"Pages 769-777"},"PeriodicalIF":3.5,"publicationDate":"2025-09-26","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"145157155","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":3,"RegionCategory":"环境科学与生态学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}

{"title":"Optimized personnel flow with minimal contamination: development and validation of an air-barrier cleanroom for cell products processing","authors":"Mitsuru Mizuno ,&nbsp;Hideaki Tani ,&nbsp;Kaori Nomura ,&nbsp;Daijiro Sone ,&nbsp;Kentaro Amano ,&nbsp;Gen Tominaga ,&nbsp;Yuki Chiba ,&nbsp;Ichiro Sekiya","doi":"10.1016/j.reth.2025.09.001","DOIUrl":"10.1016/j.reth.2025.09.001","url":null,"abstract":"<div><h3>Introduction</h3><div>Cell processing facilities for regenerative medicine require strict prevention of cross-contamination. However, the typically employed sealed, multi-room layout increases energy demands and capital costs due to heating, ventilation, and air-conditioning (HVAC), and restricts staff mobility. We devised a semi-open (SO) cleanroom that eliminated doors between the cell processing room (CPR) and adjoining corridor, while maintaining unidirectional airflow as a barrier. This study rigorously compared four interface variants—plain opening, wing walls, push–pull ventilation, and a conventional swing door—to verify whether operational flexibility can be achieved without compromising particle content performance at the CPR–corridor interface.</div></div><div><h3>Methods</h3><div>Computational fluid dynamics (CFD) simulations reproduced two connected rooms separated by a 900 × 2000-mm² opening, supplied at 23 m³/min (35 air changes per hour) constantly. Four interfaces were evaluated: plain opening, 100–500 mm wing-wall panels, push–pull ventilation adjusted to a 0.75 ratio, and a conventional swing door. A 1-m/s cross-draft emulated personnel transit. Identical full-scale mock-ups were built; particle image velocimetry (PIV) quantified airflow vectors, and optical counters logged 0.5-μm aerosols during 5-min exit and entry. The primary endpoints were the inflow particle concentration ratio across the opening and the cumulative adjacent-room transfer proportions.</div></div><div><h3>Results</h3><div>CFD showed all layouts leaked ≤0.011 %, with a 1 m/s walking draft, push–pull kept inflow below 0.05 %, halving 500-mm wing-wall performance and outperforming plain openings. The PIV confirmed significant differences in airflow velocity distributions under each condition in the case of the exit. The semi-open layout without doors showed a lower proportion of vectors pointing opposite to the forward direction than the conventional layout in both the exit and entry cases. Particle counts supported this: push–pull transferred 0.013 % of particles on exit, 32.8 % on entry, giving overall migration to the adjacent room of 0.0043 %.</div></div><div><h3>Conclusions</h3><div>The SO cleanroom concept suppresses fluctuations in particle content at the CPR–corridor interface while eliminating physical doors, enabling flexible personnel flow and obviating extra HVAC zones. Push–pull ventilation delivered the most robust containment against walking-induced disturbances, whereas the 500-mm wing walls offered a passive, power-free alternative with moderate protection. With worst-case inter-room transfers below 0.05 %, SO designs can rationally replace conventional door-sealed rooms, substantially reducing energy and construction costs in regenerative medicine manufacturing.</div></div>","PeriodicalId":20895,"journal":{"name":"Regenerative Therapy","volume":"30 ","pages":"Pages 760-768"},"PeriodicalIF":3.5,"publicationDate":"2025-09-18","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"145094918","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":3,"RegionCategory":"环境科学与生态学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}