Background Cultured oral mucosa epithelial cell sheets (COMECS) represent a promising therapeutic approach for epithelial repair, such as corneal reconstruction or esophageal mucosa defects; however, current quality assessment methods are largely invasive and destructive. Although optical flow (OF)-based analysis has shown promise in evaluating keratinocyte motility, its sensitivity decreases under confluent conditions. A novel coarse-graining (CG) analysis combined with OF is introduced as a candidate non-invasive, label-free imaging parameter to quantitatively evaluate the characteristic undulating motion patterns (local directional synchrony of cell motion) of mature COMECS and to provide a potential basis for future quality assessment. Methods COMECS were manufactured from passage 2 (p2) and passage 5 (p5) oral keratinocytes using standard (cultured in serum-containing medium) and sub-standard (cultured in serum-free medium) protocols. Time-lapse microscopy was conducted at multiple time points (48-, 96-, and 192-h post manufacture). The OF analysis-derived microscopic velocity fields were subjected to CG analysis to generate macroscopic velocity fields. The undulating index (UI) and mean undulating index (MUI) were calculated to quantify the directional synchrony of cellular motion. Western blotting was used to evaluate p63 and PITX1 protein expression levels to correlate the undulating motion patterns with biological markers of keratinocyte undifferentiation and oral epithelial identity. Results OF-based CG analysis successfully differentiated undulating motion patterns between the standard and sub-standard COMECS, with average MUI values higher in the standard COMECS (p < 0.05). Time-course analysis of p2 COMECS indicated a substantially lower average MUI at 48 h than at 96 h and 192 h, suggesting time-dependent changes in collective motion behavior. A passage-dependent comparison showed apparent temporal patterns between p2 and p5 COMECS, with p5 sheets exhibiting earlier peak undulation but reduced sustainability. The p63 and PITX1 expression levels were elevated in standard COMECS, and were associated with enhanced undulating motion patterns. Conclusion This novel OF-based, CG analysis enabled non-invasive, label-free, and quantitative assessment of COMECS quality using undulating motion pattern evaluation, demonstrating both applicability and feasibility for regenerative medicine quality control applications.
Introduction Neurogenic erectile dysfunction (ED) due to cavernous nerve injury (CNI) leads to irreversible apoptosis of the cavernosal smooth muscle and fibrosis. These structural changes are often refractory to conventional treatments. Serum-free ex vivo quality- and quantity-cultured peripheral blood mononuclear cells (MNC-QQ), which can be manufactured within 1 week from a minimal volume of peripheral blood, were developed. MNC-QQ cells are anti-inflammatory, proangiogenic, and antifibrotic, with confirmed efficacy and safety in clinical trials for refractory limb ulcers. Conditioned medium derived from MNC-QQ cells (MNCQQ-CM) contains bioactive factors with anti-inflammatory, angiogenic, and antifibrotic properties. As a cell-free, minimally invasive therapy, it represents a promising regenerative approach. In this study, we aimed to evaluate whether MNCQQ-CM promotes cavernous tissue regeneration in a CNI-induced ED model. Methods Corpus cavernosum smooth muscle cells (CCSMCs) isolated from murine penile tissue were assessed for proliferation and resistance to apoptosis following MNCQQ-CM treatment. Angiogenic capacity was evaluated through proliferation and tube formation assays using human umbilical vein endothelial cells (HUVECs). MNCQQ-CM composition was analyzed using enzyme-linked immunosorbent assay (ELISA). Bilateral CNI was induced in male rats, followed by intracavernosal injection of MNCQQ-CM, and subsequent evaluation of erectile function and histology. Results MNCQQ-CM significantly enhanced CCSMCs proliferation. Under oxidative stress, it reduced CCSMCs apoptosis. Moreover, MNCQQ-CM promoted HUVEC proliferation and tube formation, demonstrating its angiogenic potential. The presence of vascular endothelial growth factor and interleukin-10 in MNCQQ-CM was confirmed by ELISA. In CNI rats, MNCQQ-CM significantly improved erectile function and increased the smooth muscle/collagen ratio. Conclusions MNCQQ-CM demonstrates therapeutic efficacy in ED through multiple mechanisms, including angiogenesis, promotion of CCSMCs proliferation, and anti-fibrotic activity. The synergistic interaction of cytokines in MNCQQ-CM contributes to ED improvement. Given its rapid and minimally invasive production process, MNCQQ-CM holds substantial promise as a novel therapeutic agent for neurogenic diseases.
The combined application of microneedles (MNs) and exosomes represents a significant research direction in the fields of targeted drug delivery and regenerative medicine. Their synergistic and complementary effects not only address key technical challenges associated with exosome delivery—such as low delivery efficiency and limited tissue penetration—but also endow MNs with capabilities for targeted therapy and precise diagnostics, thereby demonstrating substantial advantages in the diagnosis and treatment of multisystem diseases. This review summarizes recent advances in MN-exosome delivery systems across dermatological, cardiovascular and cerebrovascular, and musculoskeletal diseases, as well as tumor diagnosis and therapy, ocular surface disorders, and oral diseases. It focuses on the fabrication strategies, mechanisms of action, and therapeutic efficacy of various MN-based exosome delivery systems. Furthermore, it proposes research perspectives and potential solutions for diseases that respond poorly to conventional diagnostic and therapeutic approaches, providing a reference for the future development and clinical translation of MN-exosome systems.
Newborn screening (NBS) is an important public health program, yet its implementation is highly uneven globally, with developing countries like Mongolia facing significant resource and infrastructure limitations that hinder early detection of preventable genetic disorders. Meanwhile, induced pluripotent stem cell (iPSC) technology, leveraged through biobanking, offers a powerful platform for disease modeling, drug discovery, and regenerative medicine, though its advanced applications are primarily concentrated in wealthier nations. We review the current status of NBS in Mongolia and argue for linking an expanded NBS program to an iPSC biobank. This link would improve diagnosis, create a foundation for Mongolian precision medicine, and strengthen local research capacity. We recognize the practical and ethical hurdles-funding, staffing, and data governance-and outline phased rollout, targeted training, and international partnerships as realistic next steps. Taken together, this approach could prevent avoidable disability in Mongolia while gradually building homegrown capability in genetic healthcare.
This commemorative article reflects on a research journey spanning neural development, stem cell biology, regenerative medicine, and iPSC-based drug discovery. My early work focused on RNA-mediated regulation in the nervous system, including studies on myelin basic protein gene regulation and the identification and functional characterization of the RNA-binding protein Musashi. These studies contributed to the conceptual foundation of neural stem cell biology and helped establish methods for identifying and isolating neural stem/progenitor cells, including those present in the adult human brain. Building on this foundation, my colleagues and I pursued translational research in spinal cord injury, ranging from analyses of injury pathophysiology and molecular interventions to preclinical studies using rodent and non-human primate models. These efforts ultimately led to the first-in-human clinical study of induced pluripotent stem cell-derived neural stem/progenitor cell transplantation for subacute spinal cord injury. In parallel, we developed patient-derived iPSC platforms for neurological disease modeling and drug discovery, particularly for amyotrophic lateral sclerosis, where iPSC-based screening identified Ropinirole as a therapeutic candidate and enabled reverse translational research linking cellular phenotypes with clinical responses. Looking ahead, I argue that the future of regenerative therapy will depend on the continued integration of developmental biology, stem cell science, disease modeling, rehabilitation, and clinical translation to address unmet medical needs in disorders of the central nervous system.
Introduction:Cell transplantation therapy using human induced pluripotent stem cell-derived cardiomyocytes (hiPSC-CMs) has shown promise for ischemic heart disease. Although paracrine effects mediated by the secretome are recognized as key mechanisms underlying the therapeutic effects of hiPSC-CMs, dynamic changes in the secretome profile during the manufacturing of hiPSC-CMs remain poorly understood. This study was aimed at elucidating the secretome dynamics of clinical-grade hiPSC-CMs to identify their "mode of action" (MoA) and "candidate quality attributes." Methods:We integrated multiomics profiling, including total RNA-sequencing, proteome analysis, and extracellular vesicle (EV)-derived microRNA-sequencing, with functional assays (cell proliferation, cell migration, and endothelial tube formation) to analyze samples obtained at different time points in the hiPSC-CM manufacturing process (Day 4-25). Results:High-purity hiPSC-CMs (>95% cardiac troponin T-positivity) exhibited distinct transcriptomic maturation between Day 16 and Day 25, characterized by the upregulation of cardiac marker genes. Proteomic clustering revealed four distinct stages, with functional transition from proliferation-centric signaling to tissue-repair signaling. Notably, the purified Day 25 secretome showed a qualitative shift toward a platelet-derived growth factor- and stromal cell-derived factor 1-rich profile. Simultaneously, EVs from the Day 25 secretome were enriched in "myomiRs" (miR-133b, -208b, -499b) and multiple anti-proliferative miRNAs (let-7e-5p, miR-145-5p). Functionally, the Day 25 secretome significantly enhanced mesenchymal stem/stromal cell (MSC) migration and promoted the formation of mature, highly branched endothelial tubes compared with the secretome from earlier stages. Conclusions:The hiPSC-CM secretome containing EVs undergoes programmed evolution during manufacturing, with functional transition from promotion of undifferentiated growth to complex tissue repair through stable angiogenesis and MSC recruitment. These findings establish a molecular foundation for the MoA of hiPSC-CM therapy and provide critical candidate quality attributes to ensure the potency and consistency of clinical-grade cardiac products.
Background:Chronic limb-threatening ischemia (CLTI) is a severe manifestation of peripheral arterial disease associated with high risks of amputation and mortality. Trials of cell-based therapies in patients unsuitable for revascularization ("no-option" or "poor-option" CLTI) have produced inconsistent results, underscoring the need for improved prognostic enrichment, baseline risk balancing, and protocol-defined stratification. Main body:This structured narrative review synthesizes evidence linking host-state markers-including laboratory biomarkers, clinical host-state factors, and physiologic/perfusion indices-to clinically meaningful CLTI outcomes. Inflammatory, nutritional, renal, metabolic, lipid, coagulation-related, infection, dialysis, frailty, smoking, etiology, and perfusion-related variables may shape the regenerative microenvironment and contribute to heterogeneity in cell therapy trial outcomes. However, current evidence primarily supports prognostic risk stratification rather than validated prediction of differential treatment response. We therefore interpret treated-cohort associations and post hoc subgroup observations as hypothesis-generating candidate variables for prospective host-state × treatment interaction testing, not as validated predictive biomarkers. We propose a conceptual trial-design framework: first, confirm no-option or poor-option CLTI through multidisciplinary review, guideline-aligned assessment, objective hemodynamic testing, vascular imaging, and multidomain assessment; second, provisionally categorize patients into favorable, intermediate, and unfavorable host-state profiles using approximate prognostic ranges for prospective validation rather than clinical eligibility cutoffs, treatment-selection rules, or gatekeeping thresholds; third, ensure clinically necessary stabilization before eligibility confirmation and treat any run-in or post-randomization optimization strategy only as an optional, secondary, protocol-justified design component; and fourth, apply stratified randomization, covariate adjustment, blinded outcome adjudication, and prespecified host-state × treatment interaction analyses. Conclusions:Integrating host-state biomarkers and clinical factors into CLTI cell therapy trial design may support prognostic enrichment, stratified enrollment, baseline risk adjustment, and hypothesis-driven evaluation of treatment-effect heterogeneity. The proposed profiles are hypothesis-generating trial-design strata, not validated clinical eligibility criteria, treatment-selection rules, or biomarker-based gatekeeping thresholds. Prospective validation will require standardized biomarker assays, harmonized endpoints, transparent reporting of cell products and concomitant care, and randomized testing of host-state × treatment interactions to advance precision regenerative medicine in this setting.
Introduction:Human amniotic membrane (HAM) is a promising scaffold for periodontal tissue engineering, supporting favorable cellular responses of human periodontal ligament fibroblasts (HPDLFs). However, molecular characterization of cells cultured on HAM is frequently hindered by extracellular matrix-derived contaminants, leading to poor RNA yield and compromised purity that limit downstream gene expression analyses. This study aims to develop and validate a matrix-adapted RNA extraction strategy to enable reliable molecular profiling of HPDLFs cultured on HAM scaffolds. Methods:HPDLFs were seeded onto de-epithelialized HAM, and cell viability was confirmed prior to RNA isolation. A systematic, multi-phase optimization of a phenol-based extraction protocol (GENEzol™ TriRNA Pure Kit) was undertaken to mitigate matrix-associated interference. RNA yield and purity were assessed spectrophotometrically, with functional validation performed using reverse transcription quantitative polymerase chain reaction (RT-qPCR). Results:The optimized protocol significantly enhanced RNA integrity and yield compared to the conventional method. Purity indices improved (A260/230: 0.73 ± 0.20 to 2.15 ± 0.40; A260/280: 1.41 ± 0.27 to 2.03 ± 0.18), while RNA concentration increased markedly. The proportion of samples meeting stringent quality criteria improved from 0% to 50%. Notably, mean RNA purity parameters were not significantly different from standard monolayer cultures for the selected measures assessed (p > 0.05). Conclusions:The approach provides a practical framework for improving RNA recovery and targeted RT-qPCR validation in HAM-based culture systems which could set precedent for improving molecular investigations in periodontal tissue engineering and other scaffold-based models.
Background:The senescence and abnormal osteogenic differentiation of bone marrow mesenchymal stem cells (BMSCs) are important causes of senile osteoporosis. Punicalagin (PUN), a natural polyphenol extracted from pomegranate peel, has demonstrated antioxidant activity in preclinical studies. This study aims to investigate the inhibitory effect of PUN on the aging of BMSCs and its therapeutic effect on senile osteoporosis. Methods:The senescence of BMSCs were induced by hydrogen peroxide (H2O2) to assess the therapeutic effect of PUN. Intracellular reactive oxygen species measurements, the senescence β-galactosidase staining, western blotting, and reverse transcription quantitative polymerase chain reaction (RT-qPCR) were conducted to evaluate the senescence of BMSCs. Alkaline phosphatase staining, Alizarin red S staining, western blotting, and RT-qPCR were performed to assess the osteogenic differentiation of senescent BMSCs. Transcriptome sequencing was conducted to explore the underlying mechanism. In vivo, the therapeutic effects of PUN were investigated in aging rats with senile osteoporosis. Results:PUN reversed the senescence of BMSCs and promoted the osteogenic differentiation in a dose-dependent manner. Mechanistically, PUN activated the nuclear factor erythroid 2-related factor 2 (NRF2)/heme oxygenase-1 (HO-1) pathway to protect BMSCs from oxidative stress. In aging rats with senile osteoporosis, intraperitoneal injection of PUN can promote osteogenesis and reduce aging-related bone loss in vivo. Conclusion:PUN alleviates senile osteoporosis by inhibiting the senescence of BMSCs via NRF2/HO-1 pathway, representing a candidate therapeutic drug for aging-related bone diseases.
Introduction:Dilated cardiomyopathy (DCM) is a major cause of heart failure with limited therapeutic options beyond heart transplantation. This study aimed to evaluated the therapeutic potential of human induced pluripotent stem cell-derived cardiomyocyte (hiPS-CM) patch transplantation in a genetic DCM model. Methods:hiPS-CM patches were transplanted onto the left ventricular surface of adult J2N-k hamsters with DCM. Cardiac function, survival, and histological evaluation were assessed. Single-nucleus RNA sequencing using left ventricular myocardium was performed to investigate cell-type-specific transcriptional alteration associated with treatment. Results:hiPS-CM patch transplantation preserved cardiac function [left ventricular ejection fraction, 41.2 (38.9 - 42.8) vs. 43.4 (40.5 - 44.6) %, P = 0.07], improved survival [Hazrod ratio 0.39 (95% confidence interval 0.16 - 0.63), P < 0.05], reduced myocardial fibrosis [fibrous area, 10.2 (9.4 - 12.1) vs. 14.1 (13.1 - 18.3) %, P < 0.05], and increased capillary density [CD-31 positive cells cout, 373.2 (336.4 - 441.3) vs. 210.7 (157.4 - 232.3) cells/mm3, P < 0.05] in J2N-k hamsters compared with controls. Transcriptomic analysis revealed regenerative shifts across multiple cell types, including activation of cardiac transcription factors (MEF2C, GATA4, NKX2-5), upregulation of angiogenesis-related pathways (ERK1/2 cascade, KLF4, ETS1, GATA2), and modulation of pro-fibrotic signaling (SMAD3, STAT3, NF-κB). Conclusions:hiPS-CM patch transplantation exerts therapeutic effects through multicellular mechanisms, including enhanced angiogenesis, improved myocardial microenvironment, and modulation of fibrosis. These findings support the feasibility of hiPS-CM patch therapy as a regenerative strategy for end-stage DCM and provide mechanistic insights into its therapeutic action.
Introduction:Environmental monitoring (EM) is essential for contamination control in regenerative medicine manufacturing, where aseptic handling and operator-dependent processes require adaptable yet robust strategies. Although EM is governed by shared regulatory requirements across manufacturing sectors in Japan, implementation varies considerably depending on product characteristics and manufacturing processes. However, quantitative investigations that position regenerative medicine within the broader landscape of regulated manufacturing sectors remain limited. This study aimed to characterize variation in EM practices across sectors in Japan and identify key factors driving these differences. Methods:A cross-sectional questionnaire survey was conducted among EM personnel at regulated pharmaceutical and regenerative medicine manufacturing facilities in Japan. Fifty-five respondents from six sectors completed a 55-item questionnaire. Fifty-four items were numerically encoded and analyzed using PCA, k-means clustering, PERMANOVA, LDA, and one-way ANOVA with effect size estimation (η2). Results:EM practices differed significantly across sectors. Major differences included implementation of surface monitoring in Grade B/C-equivalent areas, frequency and approach of media performance testing, and timing of airborne particle monitoring. Regenerative medicine facilities showed a mixed profile, combining frequent media testing with variable monitoring coverage. Clustering further separated groups based on clarity of sampling rationale and microbial identification practices. Conclusion:EM practices in regenerative medicine are partially aligned with standardized sterile manufacturing but remain heterogeneous. Identifying high-impact domains provides a basis for improving risk-based EM strategies, documentation, and training in regenerative medicine manufacturing.
Introduction:Polydactyly-derived chondrocyte (PD) sheets are being developed as an allogeneic cell sheet therapy for cartilage defects associated with knee osteoarthritis. Although intact PD sheets can promote hyaline cartilage repair, transplantation requires invasive open procedures. Injectable fragments of PD sheets (PD sheet-minis) may reduce invasiveness. However, their in vivo repair capacity after intra-articular administration remains unclear. Methods:PD cells were isolated from discarded cartilage tissue obtained during polydactyly surgery. PD sheets and PD sheet-minis were fabricated for 14 days using temperature-responsive culture inserts and RepCell plates, respectively. PD cells, PD sheets, and PD sheet-minis were compared by cell counting, viability assays, flow cytometry, soluble factor assays, and quantitative polymerase chain reaction. A full-thickness cartilage defect was created in the femoral trochlea of male athymic nude rats. The animals were randomized by body weight into vehicle, PD cell suspension, PD sheet-mini suspension, or intact PD sheet transplantation groups (n = 6 each). Hindlimb weight-bearing was assessed after surgery, and cartilage repair was evaluated histologically 4 weeks after transplantation. Results:The viable cell numbers administered per knee were comparable between PD cells, PD sheet-minis, and intact PD sheets. The PD sheet-minis and PD sheets showed similar surface marker profiles, whereas PD cells showed significantly lower CD26 and higher CD146, CD49a, and CD166 expression. Compared to PD cells, PD sheet-minis secreted more melanoma inhibitory activity and dickkopf-related protein 1 and less transforming growth factor-β, monocyte chemoattractant protein-1, and matrix metalloproteinase-3. Gene expression analysis revealed lower COL1A1 and RUNX2 expression and higher MMP3 expression in PD sheet-minis than in PD cells; COL2A1 and COL10A1 were not detected. Weight-bearing showed no sustained intergroup difference. Histologically, intact PD sheets produced consistent Safranin O-positive, type II collagen-positive, and human vimentin-positive repair tissue, whereas PD cells and PD sheet-minis mainly produced fibrous repair tissue. The mean Osteoarthritis Research Society International (OARSI) scores were 11.4 ± 5.4, 10.7 ± 6.2, 9.3 ± 7.1, and 4.4 ± 3.2 in the vehicle, PD cell, PD sheet-mini and PD sheet groups, respectively. Conclusions:PD sheet-minis retained several sheet-like in vitro characteristics; however, single intra-articular administration did not reproduce the cartilage repair achieved with intact PD sheets. These findings suggest that lesion-specific retention and persistence, rather than injectability alone, may be important determinants of structural cartilage repair by PD cell-sheet products.
Neuronal migration is a crucial process not only for brain development but also for neural regeneration after injury. It has been reported that some new neurons generated in the ventricular-subventricular zone (V-SVZ) migrate toward tissue injured by ischemic stroke and other forms of brain damage. These migrating neurons can partially compensate for lost neurons and contribute to functional recovery, including improvements in motor function. Therefore, understanding the mechanisms that regulate neuronal migration is expected to facilitate the development of novel therapeutic strategies that enhance endogenous neural regeneration after brain injury.In this review, we discuss the migratory mechanisms of new neurons generated in the V-SVZ and summarize current insights into strategies aimed at promoting neuronal migration and neuronal replacement in the injured brain.
Organoid technologies have emerged as powerful tools in regenerative medicine; however, current organoid technologies are predominantly based on pluripotent stem cell-derived systems, which poses challenges related to complexity, maturation, and clinical translation. In this commentary, we highlight a recent study by Huang et al. describing a tissue-level strategy for direct generation of functional multilineage organoids from human adult adipose tissue using reaggregated micro-fat (RMF). By preserving native tissue architecture and cellular heterogeneity, this approach enables the formation of mesoderm-, endoderm-, and ectoderm-derived organoids without genetic reprogramming or prolonged expansion culture. RMF-derived organoids demonstrate functional properties in vivo, including support of hematopoiesis and restoration of glycemic control in disease models. While further optimization and standardization will be required for clinical translation, this work provides a compelling alternative to conventional pluripotent stem cell-based paradigms and underscores the importance of tissue context in regenerative strategies. The study reframes adipose tissue as an integrated regenerative substrate and broadens the conceptual landscape of organoid generation toward more clinically accessible approaches.
Three converging therapeutic paradigms-iPSC-based drug discovery, cell transplantation, and gene therapy-have substantially expanded the therapeutic pipeline for amyotrophic lateral sclerosis (ALS) between 2020 and 2026. The FDA's accelerated approval of tofersen (Qalsody) in April 2023 marked the first treatment targeting a genetic cause of ALS. iPSC-derived drug candidates, including ropinirole and bosutinib, have completed early-phase clinical trials led by Japanese institutions. Cell therapies targeting neuroinflammation through regulatory T cells are being actively explored as immunomodulatory strategies, although efficacy remains to be established in adequately powered trials. Next-generation gene-silencing approaches-including RNA interference (RNAi) therapeutics and AAV-delivered microRNA-entered first-in-human trials in 2024-2025. The identification of STMN2 as a downstream target of TDP-43 dysfunction has opened a potential TDP-43-downstream nucleic acid therapeutic avenue for sporadic ALS, which constitutes approximately 90% of all cases, with company-reported interim data suggesting target engagement in the ongoing Phase 1/2 ANQUR trial (QRL-201). This review synthesizes the latest evidence across all three therapeutic domains, with attention to the hierarchy of evidence, regulatory milestones, and the pioneering contributions of Japanese research groups.
Introduction:Lung injury is a frequent complication following heart failure (HF) due to systemic inflammation, hypoxia, and reduced cardiac function. This study compared the therapeutic effects of menstrual blood-derived stromal cells (MenSCs) and bone marrow-derived stromal cells (BMSCs) on HF-induced lung injury in rats. Methods:Twenty male Wistar rats underwent permanent ligation of the left anterior descending coronary artery to induce myocardial infarction (MI). They were randomly assigned to receive intravenous injections of MenSCs (2 × 106 cells), BMSCs (2 × 106 cells), or PBS 7 days post-MI. Cardiac function was assessed by echocardiography, and lung tissues were analyzed 28 days post-cell administration. Histopathology (H&E and Masson's trichrome staining) evaluated pulmonary structural changes and fibrosis, TUNEL assay assessed apoptosis, and immunohistochemistry measured NF-κB and iNOS expression. Results:Results showed that MenSC treatment significantly improved left ventricular ejection fraction compared with MI and BMSC groups (both P < 0.0001). MenSCs markedly reduced pulmonary injury scores and apoptotic cell numbers compared with both untreated and BMSC-treated rats (P < 0.0001). Both cell types reduced pulmonary fibrosis compared to MI (P < 0.0001). Both MenSCs and BMSCs similarly reduced NF-κB expression, whereas only MenSCs effectively decreased iNOS expression (P < 0.0001). Human mitochondrial antibody staining confirmed successful homing of both cell types to injured lung tissue. Conclusion:systemic administration of MenSCs effectively ameliorated HF-induced lung injury through anti-inflammatory, anti-apoptotic, and anti-fibrotic mechanisms. Compared with BMSCs, MenSCs demonstrated superior therapeutic potential, highlighting their promise as a non-invasive and immunologically favorable cell source for regenerative therapy in remote organ injury following MI.
Introduction:Reliable surface disinfection of biosafety cabinets is essential to prevent contamination in pharmaceutical, clinical, and laboratory environments. However, manual cleaning using ethanol (ETH) and wiping is subject to operator variability, making objective verification of disinfection quality difficult. This paper proposes an integrated disinfection-validation system that combines thermal imaging, visual tracking, and pressure sensing to objectively assess the adequacy of ETH spraying and pressure-based wiping. Methods:The proposed system integrated three sensing modalities: Red-green-blue (RGB) imaging, thermal sensing, and pressure detection. The RGB and thermal cameras (placed outside the biosafety cabinet) monitored the stainless steel (SUS) work surface, and the flexible pressure sensors (embedded in the wiping tool) detected the applied force. The system was biologically validated using Bacillus subtilis spores on SUS plates, using ETH spraying only, wiping only, and combined spraying and pressure-based wiping. Reproducibility was evaluated under different wiping speeds and operator techniques, using time-series and endpoint analyses. Results:The system successfully visualized the sprayed, wiped, and untreated areas in real time. Only the combined treatment resulted in complete inhibition of bacterial growth, demonstrating the synergistic effect of chemical and mechanical actions in achieving effective disinfection. The classification accuracy and quantitative detection of the cleaned areas were consistent across the trials, confirming their reproducibility under variable wiping patterns. Conclusions:This integrated sensing system provides a reliable and objective method for validating surface disinfection in biosafety cabinets. By enabling the real-time, recordable, and quantitative assessment of manual cleaning, it bridges the gap between procedural operations and verifiable decontamination, supporting standardized and data-driven hygiene management in biosafety and cell processing environments.
Introduction:Primary chondrocytes often lose matrix-forming capacity during in vitro expansion, limiting scalable cartilage engineering. Here, we examined aging-independent regulation of chondrogenic function during long-term expansion of primary auricular chondrocytes and sought molecular features associated with late-passage functional decline. Methods:Mouse auricular chondrocytes (mACs) were serially expanded in 2D culture up to passage 40 or 50 (P50) under high- or low-seeding conditions. Proliferation, morphology, and senescence status were assessed by growth curves, phase-contrast imaging, and SA-β-gal staining. Chondrogenic capacity was evaluated using 3D pellet culture and toluidine blue staining. Bulk RNA sequencing of matched 2D and 3D samples was followed by PCA, differential expression, and pathway enrichment analyses. Key transcriptional signatures were validated by RT-qPCR in mouse auricular chondrocytes. Results:Both seeding conditions supported sustained proliferation across extended passaging. Although not immortalized, mACs maintained stable proliferation for more than P 50 with minimal senescence-associated features, exhibiting cell line-like proliferative behavior. Despite preserved growth, proteoglycan-rich matrix production declined at P50. Transcriptomic analyses identified culture dimensionality as the dominant source of variance, with 2D cultures showing passage-dependent drift and 3D pellets retaining a more stable chondrogenic profile. Cross-comparisons identified a conserved nine-gene decline module (Gpx7, Krba1, Plxdc2, Ptgr2, Rarres1, Pxdc1, Ctsh, Pigh, and Diras2), which was validated by RT-qPCR. Enrichment analyses indicated coordinated attenuation of adhesion-, extracellular-, and differentiation-related programs at late passages. Conclusion:Auricular chondrocytes exhibit cell line-like proliferative capacity with limited senescence activation while progressively losing matrix-forming competence at ultra-late passages. A conserved nine-gene decline module provides molecular benchmarks for passage-resolved assessment and may inform optimization of scalable auricular cartilage manufacturing.
