Regenerative Therapy最新文献

{"title":"Role of WIF-1 in regulating chondrocyte dedifferentiation and matrix production through WNT pathway","authors":"Yu-Ying Chu ,&nbsp;Yukiyo Asawa ,&nbsp;Naoki Tsuji ,&nbsp;Tomoaki Sakamoto ,&nbsp;Dan Liu ,&nbsp;Kazuto Hoshi ,&nbsp;Atsuhiko Hikita","doi":"10.1016/j.reth.2026.101083","DOIUrl":"10.1016/j.reth.2026.101083","url":null,"abstract":"<div><h3>Introduction</h3><div>Chondrocyte dedifferentiation during monolayer expansion is a major barrier to producing clinically reliable cells for cartilage regeneration. The upstream regulators governing this phenotypic instability are not fully defined. This study identifies WNT inhibitory factor 1 (WIF1) as a potential key molecular determinant of chondrocyte stability.</div></div><div><h3>Methods</h3><div>Human auricular chondrocytes (HACs) were expanded to early (P2) and late (P7) passages. Transcriptomic profiling, qPCR, GAG quantification, and histology were performed in 2D monolayer and 3D atelocollagen pellet cultures. WIF1 function was interrogated using lentiviral knockdown and overexpression. β-catenin, <em>p</em>-SMAD2/3, <em>p</em>-JNK, COL1A1, COL2A1, and WIF1 were evaluated by immunohistochemistry.</div></div><div><h3>Results</h3><div>Late-passage HACs showed marked dedifferentiation, including reduced WIF1, COL2A1, ACAN, and SOX9 with concomitant increases in COL1A1, ACTA2, and MMP9. WIF1 knockdown in early-passage cells reproduced this phenotype, inducing β-catenin nuclear accumulation, increased SMAD2/3 and JNK activation, diminished GAG synthesis, and loss of cartilage-specific ECM architecture. Conversely, WIF1 overexpression in P7 chondrocytes restored chondrogenic markers, enhanced proteoglycan-rich ECM formation, and suppressed fibroblastic gene expression. WIF1 overexpression re-established a signaling profile resembling early-passage cells, characterized by low β-catenin, reduced <em>p</em>-SMAD2/3, and attenuated <em>p</em>-JNK.</div></div><div><h3>Conclusions</h3><div>WIF1 functions as a critical upstream regulator that maintains chondrocyte identity by simultaneously suppressing WNT/β-catenin, TGF-β/SMAD, and JNK stress signaling. Its loss is a hallmark of dedifferentiation, while its restoration reverses transcriptional drift and rescues ECM-producing capacity. These findings highlight WIF1 as a promising therapeutic target to improve the stability of expanded chondrocytes and enhance the quality of regenerative cartilage constructs.</div></div>","PeriodicalId":20895,"journal":{"name":"Regenerative Therapy","volume":"31 ","pages":"Article 101083"},"PeriodicalIF":3.5,"publicationDate":"2026-03-01","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"147420278","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":"Efficacy and safety of autologous adipose-derived stem cells combined with platelet-rich plasma for periodontal regeneration: a multicenter, randomized, open-label, parallel-group comparative trial","authors":"Morikuni Tobita ,&nbsp;Yosuke Masubuchi ,&nbsp;Yorimasa Ogata ,&nbsp;Akio Mitani ,&nbsp;Takeshi Kikuchi ,&nbsp;Jun-Ichiro Hayashi ,&nbsp;Taku Toriumi ,&nbsp;Anna Arita ,&nbsp;Jorge Luis Montenegro Raudales ,&nbsp;Hiroshi Mizuno ,&nbsp;Yuki Suzuki ,&nbsp;Keiko Wakana ,&nbsp;Hikari Yoneda ,&nbsp;Masahiro Kino-oka ,&nbsp;Tomohiro Morio ,&nbsp;Kiyoshi Okada ,&nbsp;Shinya Murakami ,&nbsp;Masaki Honda","doi":"10.1016/j.reth.2026.101062","DOIUrl":"10.1016/j.reth.2026.101062","url":null,"abstract":"<div><h3>Introduction</h3><div>Periodontitis is a chronic inflammatory disease caused by dental biofilm that destroys the periodontal tissues supporting the teeth. We aimed to evaluate the efficacy and safety of co-transplantation of autologous adipose-derived mesenchymal stem cells (ASCs) with platelet-rich plasma (PRP) compared to enamel matrix derivative (EMD) for regenerating periodontal tissue damaged by chronic periodontitis.</div></div><div><h3>Methods</h3><div>In this multicenter, randomized, open-label, parallel-group phase II therapeutic equivalence trial, we assessed the effects of ASCs with PRP versus EMD on alveolar bone height and width, as well as other periodontal indices. Measurements of primary and secondary endpoints and periodontal tissue indices were performed at baseline, the day of transplantation, and at 12, 24, and 36 weeks post-transplantation. Data were analyzed using Student's t-test.</div></div><div><h3>Results</h3><div>Of the 21 patients initially recruited, 9 in the ASCs + PRP group and 6 in the EMD group completed the study. The ASCs + PRP treatment demonstrated greater regenerative potential than that of EMD, with a significantly higher alveolar bone height at 36 weeks post-treatment. Patients receiving ASCs + PRP treatment showed a statistically higher mean height of new alveolar bone in the transplanted area than those in the EMD group. Clinical attachment levels improved significantly from baseline at 12, 24, and 36 weeks in the ASCs + PRP group; however, no significant difference in clinical attachment gain was observed between the two groups. Adverse events in the ASCs + PRP group were neither related to the implantation site nor to the treatment.</div></div><div><h3>Conclusions</h3><div>Co-transplantation of ASCs with PRP is a safe and effective approach for periodontal regeneration, offering a promising therapeutic strategy for the treatment of periodontitis.</div></div>","PeriodicalId":20895,"journal":{"name":"Regenerative Therapy","volume":"31 ","pages":"Article 101062"},"PeriodicalIF":3.5,"publicationDate":"2026-03-01","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"145976703","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":"Extracellular vesicles derived from adipose-derived mesenchymal stem/stromal cells prevent synovial inflammation and attenuate cartilage degeneration in rodent osteoarthritis","authors":"Yushi Maruiwa ,&nbsp;Yasuo Niki ,&nbsp;Yo Mabuchi ,&nbsp;Osamu Takeuchi ,&nbsp;Satoshi Kuronuma ,&nbsp;Yoshitsugu Fukuda ,&nbsp;Atsuhiro Fujie ,&nbsp;Akihito Oya ,&nbsp;Shu Kobayashi ,&nbsp;Masaya Nakamura","doi":"10.1016/j.reth.2025.101056","DOIUrl":"10.1016/j.reth.2025.101056","url":null,"abstract":"<div><h3>Objective</h3><div>Osteoarthritis (OA) is the most common joint disease in the elderly and a major cause of pain and disability. Recent advances in OA therapy have led to a greater variety of treatment options. Extracellular vesicles (EVs) have recently emerged as a potential therapeutic approach for OA. This study aimed to demonstrate the disease-modifying effects of adipose-derived mesenchymal stem cell-derived EVs (MSC-EVs) on OA.</div></div><div><h3>Methods</h3><div>The anti-inflammatory effects of MSC-EVs were evaluated using RAW264.7 macrophage-like cells stimulated with lipopolysaccharide and human synovial cells stimulated with IL-1β. In vivo, MSC-EVs were administered intra-articularly into knees of rats with monosodium iodoacetate (MIA)-induced OA. Pain thresholds, gait parameters, histological scores, and cytokine levels were assessed. Single-cell RNA sequencing (scRNA-seq) was performed on joint tissues to evaluate cell-specific gene expression and macrophage polarization.</div></div><div><h3>Results</h3><div>In vitro, MSC-EVs reduced the mRNA expression of IL-1β and IL-6 in RAW264.7 cells, and significantly suppressed multiple inflammatory cytokines while upregulating FGF-18 in synovial cells. In vivo, intra-articular MSC-EV injection increased pain threshold, improved gait, and reduced synovial inflammation and cartilage degeneration. scRNA-seq revealed decreased inflammatory cytokines, increased PRG4 and FGF-18 expression, and a shift in macrophage polarization toward an M2 phenotype in the EV-treated group.</div></div><div><h3>Conclusion</h3><div>MSC-EVs exert anti-inflammatory and cartilage-protective effects in OA through immune modulation and regenerative signaling, indicating their significant therapeutic potential as a disease-modifying strategy for OA.</div></div>","PeriodicalId":20895,"journal":{"name":"Regenerative Therapy","volume":"31 ","pages":"Article 101056"},"PeriodicalIF":3.5,"publicationDate":"2026-03-01","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"145883644","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":"First-in-human study to investigate the safety and efficacy of effective-mononuclear cell therapy for radiogenic xerostomia","authors":"Takashi I ,&nbsp;Mayumi Iwatake ,&nbsp;Takako Yoshida ,&nbsp;Kazuhiro Nagai ,&nbsp;Yuka Hotokezaka ,&nbsp;Mika Nishihara ,&nbsp;Ryo Honma ,&nbsp;Hiroshi Harada ,&nbsp;Munehiro Hayashida ,&nbsp;Toshiki Nagano ,&nbsp;Riho Kanai ,&nbsp;Seigo Ohba ,&nbsp;Atsutoshi Yoshimura ,&nbsp;Haruchika Masuda ,&nbsp;Takayuki Asahara ,&nbsp;Yasushi Miyazaki ,&nbsp;Atsushi Kawakami ,&nbsp;Izumi Asahina ,&nbsp;Makoto Seki ,&nbsp;Yoshinori Sumita","doi":"10.1016/j.reth.2025.101059","DOIUrl":"10.1016/j.reth.2025.101059","url":null,"abstract":"<div><h3>Background</h3><div>Salivary gland (SG) hypofunction is the most common complication following radiotherapy for head and neck cancer. Currently, there are no effective therapies for radiation-induced xerostomia. We recently developed a novel therapy for preclinical studies using effective-mononuclear cells (E-MNCs) induced from autologous peripheral blood mononuclear cells (PB-MNCs) via a new primary culture system for treatment of radiation-induced xerostomia. However, the safety and effectiveness of E-MNC therapy have not been assessed in humans. The objective of this first-in-human study was to evaluate the safety, tolerability, and in part the efficacy of E-MNC therapy for treating radiation-induced xerostomia.</div></div><div><h3>Methods</h3><div>This first-in-human study was an open-label, single-center, two-step dose escalation study. A total of 5 patients who had no recurrence of head and neck cancer over a 5-year period following radiation therapy and suffered from radiation-induced xerostomia received a transplantation of E-MNCs to one-sided submandibular gland (SMG). The primary endpoint was the safety of the protocol. The secondary endpoint was effectiveness evaluated based on change from baseline in whole-saliva secretion, MRI/CT-evaluated volume of the SMG and subjective/objective symptoms after intervention. The duration of the intervention was 1 year.</div></div><div><h3>Results</h3><div>During follow-up, no treatment-related adverse events were detected. The stimulated whole salivary flow rate increased from an average of 3.86 mL/10 min at baseline to 5.16 mL/10 min at 6 months post–investigational intervention. Additionally, subjective/objective symptoms improved in 4 of 5 treated patients. By contrast, imaging findings revealed no obvious changes in MRI/CT-evaluated volume of SMGs due to conspicuous progression of atrophy and fibrosis compared with baseline.</div></div><div><h3>Conclusion</h3><div>This is the first clinical study to evaluate the safety and efficacy of E-MNC treatment in patients with severe radiation-induced xerostomia. The results of our study indicate that E-MNC treatment is safe and effective, and provide valuable information that will aid in designing subsequent clinical studies.</div></div><div><h3>Trial registration</h3><div>This study was registered with the Japan Registry of Clinical Trials (<span><span>http://jrct.niph.go.jp</span><svg><path></path></svg></span>) as jRCTb070190057.</div></div>","PeriodicalId":20895,"journal":{"name":"Regenerative Therapy","volume":"31 ","pages":"Article 101059"},"PeriodicalIF":3.5,"publicationDate":"2026-03-01","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"145924579","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":"Immune synapse molecules knockout in induced pluripotent stem cells enables broad NK cell resistance in T cell progeny","authors":"Jing Zhang ,&nbsp;Keisuke Sumide ,&nbsp;Keitaro Kanie ,&nbsp;Akihiro Ishikawa ,&nbsp;Munehiro Yoshida ,&nbsp;Tomoko Ishii ,&nbsp;Bo Wang ,&nbsp;Shin Kaneko","doi":"10.1016/j.reth.2026.101060","DOIUrl":"10.1016/j.reth.2026.101060","url":null,"abstract":"<div><h3>Introduction</h3><div>Allogeneic iPSCs provide a potential source for regenerative T cell therapies, yet their clinical application remains limited by immune rejection driven by human leukocyte antigen (HLA) incompatibility. Knockout of β2-microglobulin (B2M) eliminates HLA class I expression and protects against CD8⁺ T cell-mediated killing, but paradoxically provokes natural killer (NK) cell activation via the “missing-self” response. Existing approaches seek to enhance inhibitory signaling or dampen activating signals, yet these approaches achieve only partial NK evasion due to the extraordinary heterogeneity of NK receptor repertoires.</div></div><div><h3>Methods</h3><div>We developed a broader and more universal immune evasion strategy by targeting immune synapse (IS) adhesion molecules that critical for NK–target cell engagement. Building on our previously reported hypoimmunogenic iPSC-derived T cell (iT cell) platform (B2M^KOCIITA^KOPVR^KO with HLA-E overexpression), we engineered double-knockout (dKO) iT cells lacking CD54 (ICAM-1) and CD58 (LFA-3) using CRISPR/Cas9-mediated gene editing. These two ligands play key roles in stabilizing NK–target immunological synapses.</div></div><div><h3>Results</h3><div>Functionally, dKO iT cells exhibited marked resistance to NK cell–mediated cytotoxicity both in vitro and in vivo in human IL-15 transgenic mouse model, while maintaining iT cell cytotoxic effector functions. By disrupting the physical interface required for NK engagement, this approach provides broad protection against diverse NK cell subsets and complements existing HLA-focused immune evasion strategies.</div></div><div><h3>Conclusions</h3><div>Our findings establish a potentially versatile platform for generating universal, NK-resistant iT cells and advance the translational potential of iPSC-based immunotherapies.</div></div>","PeriodicalId":20895,"journal":{"name":"Regenerative Therapy","volume":"31 ","pages":"Article 101060"},"PeriodicalIF":3.5,"publicationDate":"2026-03-01","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"145976704","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":"Stable and functional regulatory T cell attenuates fibrotic remodeling in heart failure","authors":"Takura Taguchi ,&nbsp;Takuji Kawamura ,&nbsp;Norihisa Mikami ,&nbsp;Kosuke Torigata ,&nbsp;Akima Harada ,&nbsp;Lisa Fujimura ,&nbsp;Emiko Ito ,&nbsp;Maki Takeda ,&nbsp;Kenji Miki ,&nbsp;Shimon Sakaguchi ,&nbsp;Shigeru Miyagawa","doi":"10.1016/j.reth.2026.101079","DOIUrl":"10.1016/j.reth.2026.101079","url":null,"abstract":"<div><h3>Introduction</h3><div>Heart failure involves myocardial fibrosis and impaired cardiac function. The transverse aortic constriction (TAC) model in mice replicates pressure overload-induced fibrosis. Immune-mediated inflammation, particularly through T lymphocytes and macrophages, plays a pivotal role in the progression of fibrosis. In vitro–induced regulatory T cells (Tregs) are limited by Foxp3 instability. To address this, stable and functional in vitro induced Tregs (S/F-iTregs) have been developed under CD28-deprived conditions, exhibiting sustained Foxp3 expression and epigenetic stability. We aimed to evaluate the therapeutic efficacy of intravenously administered S/F-iTregs in a murine TAC model of heart failure.</div></div><div><h3>Methods</h3><div>S/F-iTregs were generated from CD4⁺ conventional T cells under CD28 costimulation-deficient conditions. BALB/c mice underwent TAC surgery and received S/F-iTregs intravenously. Cardiac function was assessed by echocardiography. Myocardial fibrosis was evaluated using histological staining. Bulk RNA sequencing was performed to assess transcriptomic changes in the myocardium.</div></div><div><h3>Results</h3><div>S/F-iTreg administration significantly improved left ventricular function and reduced myocardial fibrosis compared to those in controls. PKH26-labeled S/F-iTregs were detected within the cardiac tissue, confirming in vivo migration. RNA sequencing revealed downregulation of inflammation- and fibrosis-related pathways, including tumor necrosis factor-alpha/nuclear factor kappa-light-chain-enhancer of activated B cell, platelet-derived growth factor, transforming growth factor-beta signaling, and modulation of macrophage-related transcriptional programs.</div></div><div><h3>Conclusions</h3><div>Systemic administration of S/F-iTregs attenuates cardiac fibrosis and preserves cardiac function in pressure overload–induced heart failure. These findings support S/F-iTreg–based immunomodulation as a potential approach to limit adverse fibrotic cardiac remodeling.</div></div>","PeriodicalId":20895,"journal":{"name":"Regenerative Therapy","volume":"31 ","pages":"Article 101079"},"PeriodicalIF":3.5,"publicationDate":"2026-03-01","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"147378372","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":"Intra-articular injections of platelet-rich plasma suppress cartilage degeneration in a mouse model of knee osteoarthritis","authors":"Yasumasa Momoi ,&nbsp;Yoshitomo Saita ,&nbsp;Ryosuke Nakajima ,&nbsp;Sayuri Uchino ,&nbsp;Hirofumi Nishio ,&nbsp;Shin Fukusato ,&nbsp;Takanori Wakayama ,&nbsp;Yohei Kobayashi ,&nbsp;Atsushi Furuhata ,&nbsp;Hiroshi Ikeda ,&nbsp;Kazuo Kaneko ,&nbsp;Muneaki Ishijima","doi":"10.1016/j.reth.2026.101072","DOIUrl":"10.1016/j.reth.2026.101072","url":null,"abstract":"<div><h3>Introduction</h3><div>Pain-relieving drugs, such as nonsteroidal anti-inflammatory drugs and hyaluronic acid, are standard treatments for knee osteoarthritis (OA); however, no disease-modifying drugs exist for knee OA. Platelet-rich plasma (PRP) is a novel treatment with both symptom improvement and disease-modifying effects; however, the underlying mechanism remains unknown. In addition, the biologically active substances contained in PRP differ greatly depending on the purification method used. This controlled laboratory study aimed to investigate the therapeutic effects of two types of PRP with different white blood cell concentrations on knee OA using an animal model.</div></div><div><h3>Methods</h3><div>Leukocyte-rich PRP (LR-PRP) and leukocyte-poor PRP (LP-PRP) were prepared from 10-week-old female C57BL/6 mice. A mouse model of knee osteoarthritis was generated by the unilateral transection of the medial meniscus in the right hind limb. Mice were randomly assigned to three treatment groups that received 6 μL intra-articular injections of either phosphate-buffered saline (control), LR-PRP, or LP-PRP at 2-, 4-, and 6-weeks post-surgery. Mice were sacrificed 12 weeks post-surgery and histologic analysis, immunohistochemistry analysis for CD68 and three‐dimensional micro-computed tomography (3DμCT) of knee joints were analyzed. Hind limb weight-bearing distribution was measured preoperatively and at 4- and 12-weeks post-surgery. Statistical analyses were performed using GraphPad Prism 9.0.2. P-values of &lt;5 % were considered statistically significant.</div></div><div><h3>Results</h3><div>Histological analysis of the femoral medial condyle (Osteoarthritis Research Society International (OARSI) score) showed that both the LP and LR groups had significantly suppressed cartilage destruction compared with the Phosphate Buffered Saline (PBS) group (P = 0.01). The percentage of CD68-positive synovial macrophages in the lateral joint was significantly lower in the LR group than in the PBS group (PBS: 2.0 ± 2.9 %, LR: 0.6 ± 1.3 %, LP: 0.9 ± 1.9 %; P = 0.02). The affected-side load rate (%) increased in the LR and LP groups, with a significant increase observed in the LR group from week four. PBS group, Pre/4w/12w = 36.0 ± 4.8/33.8 ± 6.8/36.2 ± 7.4 % (P = 0.60); LR group, Pre/4w/12w = 31.9 ± 2.2/34.3 ± 8.3/44.2 ± 4.8 % (P &lt; 0.01); LP group, Pre/4w/12w = 33.5 ± 7.4/36.1 ± 7.4/42.0 ± 5.2 % (P = 0.02). Conversely, no significant difference in BMD was observed between groups.</div></div><div><h3>Conclusions</h3><div>Intra-articular injection of LR- and LP-PRP attenuated cartilage degeneration in the medial femoral condyle in a mouse model of knee osteoarthritis.</div></div>","PeriodicalId":20895,"journal":{"name":"Regenerative Therapy","volume":"31 ","pages":"Article 101072"},"PeriodicalIF":3.5,"publicationDate":"2026-03-01","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"146077152","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":"Butyrate-preconditioned human adipose-derived stem cell-conditioned medium enhances myocardial perfusion after infarction","authors":"Shinji Kobuchi ,&nbsp;Wan-Tseng Hsu ,&nbsp;Misaki Matsuzawa ,&nbsp;Rina Kagawa ,&nbsp;Junko Watanabe ,&nbsp;Koki Harada ,&nbsp;Yuki Toda ,&nbsp;Shohei Hamada ,&nbsp;Masayuki Tsujimoto ,&nbsp;Hidekazu Kawashima ,&nbsp;Kaneyasu Nishimura ,&nbsp;Kenjiro Matsumoto ,&nbsp;Kazuyuki Takata","doi":"10.1016/j.reth.2025.11.007","DOIUrl":"10.1016/j.reth.2025.11.007","url":null,"abstract":"<div><h3>Introduction</h3><div>Adipose-derived stem cell-conditioned medium (ASC-CM) is promising for cardiac repair via paracrine mechanisms. However, variability in efficacy limits its clinical translation. We investigated whether preconditioning human ASC with butyrate (ASC-BA-CM) enhanced its paracrine potency and improved <em>in vitro</em> and <em>in vivo</em> outcomes.</div></div><div><h3>Method</h3><div>RNA-sequencing of human ASCs treated with butyrate was performed to characterize transcriptomic changes. CM was collected and analyzed via cytokine/chemokine arrays. Wound healing assays using human umbilical vein endothelial cells (HUVECs), with and without THP-1 macrophage co-culture, were performed to evaluate endothelial repair and its correlations with secreted factors. <em>In vivo</em> angiogenesis was assessed using a sponge implantation model, and myocardial perfusion was measured in a rat myocardial infarction model using single-photon emission computed tomography/computed tomography (SPECT/CT) thallium-201 imaging.</div></div><div><h3>Results</h3><div>Butyrate preconditioning upregulated angiogenesis- and immune-related genes, including <em>CXCL8</em>, <em>SOD2</em>, and <em>TGM2</em>. It increased IL-10, CXCL5, and MMP-1 secretion. <em>In vitro</em>, BA-preconditioned ASC-CM enhanced HUVEC wound closure, which was improved by co-culture with THP-1 macrophages and negatively correlated with TGFb3 and TIMP-2 levels. <em>In vivo</em>, ASC-BA-CM promoted vascularization and macrophage accumulation in sponges and significantly improved myocardial perfusion by approximately 32 % compared with controls.</div></div><div><h3>Conclusions</h3><div>Butyrate preconditioning enhanced the paracrine activity of ASC-CM and was associated with improved myocardial perfusion in a rat model. These findings suggest that butyrate may augment the ASC secretome function. Potential mechanisms such as endothelial repair, angiogenesis, and immune modulation remain hypothetical and require further validation in future studies.</div></div>","PeriodicalId":20895,"journal":{"name":"Regenerative Therapy","volume":"31 ","pages":"Article 101042"},"PeriodicalIF":3.5,"publicationDate":"2026-03-01","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"145578352","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":"Influence of donor–recipient age differences in allogenic rat thyroid transplantation","authors":"Ayumi Arauchi ,&nbsp;Katsuhisa Matsuura ,&nbsp;Tatsuya Shimizu","doi":"10.1016/j.reth.2025.101047","DOIUrl":"10.1016/j.reth.2025.101047","url":null,"abstract":"<div><h3>Introduction</h3><div>Regenerative medicine for tissue dysplasia, hypoplasia, and functional impairment of tissues and cells has advanced considerably, and thyroid disease is no exception. Inducing differentiation of thyroid cells from patient-derived cells and transplanting them back into patients is an ideal approach because it eliminates the need for immunosuppressive drugs. However, the relationship between the maturity of cells or tissues derived from various sources and engraftment outcomes remains unclear. In this study, we evaluated the effect of donor–recipient age differences using thyroids from living rat donors, given the current lack of sufficiently mature stem cell-derived thyroid tissue.</div></div><div><h3>Methods</h3><div>Histological and gene expression differences were analyzed in thyroids of retired rats (&gt;20 weeks old with impaired reproductive function) and 3-week-old rats. Thyroid transplantation experiments were conducted between age-matched or age-mismatched groups. Donor thyroids were implanted beneath the renal capsule of recipient rats without total thyroidectomy, and grafts from retired rats transplanted into 3-week-old rats were resected 1 and 6 months post-transplantation, while others were resected 1 month post-transplantation for immunohistological analysis and gene expression analysis of thyroid differentiation markers.</div></div><div><h3>Results</h3><div>Thyroids from 3-week-old rats and retired rats showed minimal histological and functional differences; however, the expression levels of several thyroid-specific marker genes were significantly higher in retired rats. When thyroids were transplanted between age-matched donors and recipients, clear engraftment was observed at 1 month. Although robust engraftment was also observed when thyroids from retired rats were transplanted into 3-week-old recipients at both 1 and 6 months, transplantation of thyroids from 3-week-old donors into retired rats resulted in disrupted follicular structures and fibrotic changes at 1 month. In contrast, mRNA expression levels of transplanted thyroids presented no significant differences between age-matched and age-mismatched transplantation.</div></div><div><h3>Conclusions</h3><div>Mismatch between donor thyroid maturity and recipient age may influence engraftment in rat allogenic thyroid transplantation.</div></div>","PeriodicalId":20895,"journal":{"name":"Regenerative Therapy","volume":"31 ","pages":"Article 101047"},"PeriodicalIF":3.5,"publicationDate":"2026-03-01","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"145691074","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":"Development of a digital analysis system for a novel 3D culture-based colony formation to detect malignantly transformed cells in human cell-based therapeutic products","authors":"Shinji Kusakawa ,&nbsp;Tingshu Yang ,&nbsp;Rumi Sawada ,&nbsp;Ryuji Kato ,&nbsp;Yoji Sato ,&nbsp;Satoshi Yasuda","doi":"10.1016/j.reth.2025.101052","DOIUrl":"10.1016/j.reth.2025.101052","url":null,"abstract":"<div><h3>Introduction</h3><div>The presence of malignantly transformed cells in human cell-based therapeutic products (hCTPs) is a significant safety concern. Although such cellular impurities in hCTPs can be assessed by detecting anchorage-independent growth using conventional soft agar colony formation (SACF) assays, the sensitivity of these assays is often insufficient. To overcome this limitation, we previously developed a novel tumorigenicity-associated testing method, the digital SACF (D-SACF) assay, which combines a partitioned culture of test cells to concentrate target cells with colony detection via image analysis. However, conventional soft agar culture involves complicated operations, such as preparing multilayered agar media and temperature control, and further technical optimization is required for the widespread adoption of the D-SACF assay.</div></div><div><h3>Methods</h3><div>In this study, we focused on a new culture system incorporating a three-dimensional (3D) culture method using a liquid medium containing the low-molecular-weight agar polymer LA717 in low-adhesion culture vessels. We initially confirmed conditions for the efficient high-density 3D culture of normal cells using LA717-supplemented medium in low-adhesion 96-well plates.</div></div><div><h3>Results</h3><div>Using human mesenchymal stem/stromal cells (MSCs) as a normal cell model and HeLa cells as a transformed cell model, we demonstrated that the new 3D culture system effectively maintained the dispersion of MSCs and prevented their aggregation, while transformed HeLa cells exhibited robust anchorage independence, thereby establishing the new liquid/low-molecular-weight agar colony formation (LACF) method as an alternative to SACF.</div></div><div><h3>Conclusions</h3><div>Finally, by systematizing the digital analysis system for the LACF assay (D-LACF assay), which streamlines the overall workflow from the performance evaluation of the test method to product testing and result interpretation, the limitations of the conventional soft agar-based D-SACF assay were addressed, and its practicality and utility were enhanced. This <em>in vitro</em> evaluation system is expected to provide a promising approach for improving the quality and safety of hCTPs.</div></div>","PeriodicalId":20895,"journal":{"name":"Regenerative Therapy","volume":"31 ","pages":"Article 101052"},"PeriodicalIF":3.5,"publicationDate":"2026-03-01","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"145796775","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}