Premature ovarian insufficiency (POI) is defined by ovarian dysfunction and consequent decreased fertility. While follicular depletion is an acknowledged factor, dynamic changes in follicular subpopulations and single-cell-level niche remodeling remains largely unexplored. Human amniotic epithelial cells (hAECs) represent a promising regenerative approach for restoring ovarian function; however, the underlying mechanism in promoting follicular development remains unclear. We established a chemotherapy-induced POI mouse model and conducted single-nucleus RNA sequencing (snRNA-seq) to systematically characterize ovarian cellular heterogeneity, residual follicular development, and ovarian microenvironmental alteration. A human-derived cell projection model was established to track the distribution of transplanted hAECs. Furthermore, the functional role of IGF1R signaling in granulosa cells was validated using both in vitro culture assays and in vivo interventions. We identified eight distinct ovarian cell types and uncovered stage-specific injury patterns: acute chemotherapy exposure induced massive loss of granulosa cells and oocytes, while the chronic phase was characterized by fibrotic accumulation and immune infiltration. Furthermore, residual follicles exhibited aberrant development trajectories and compromised progression, primarily due to disrupted granulosa cells -oocytes communication. Notably, hAEC transplantation significantly enhanced follicle survival, partially attenuated fibrosis and promoted ovarian structural restoration. Mechanistically, hAECs-derived IGFBP2 regulated IGF1R expression in granulosa cells, thereby reactivating the Akt/FoxO3A signaling pathway and downregulating the senescence marker P21. This study presents a time-resolved snRNA-seq atlas capturing both acute and chronic injury phases in a chemotherapy-induced POI model, combined with a human-mouse projection approach to track transplanted hAECs in damaged ovaries. Our study establishes a novel cell-based therapeutic strategy for partial ovarian functional recovery, in which hAEC-derived IGFBP2 restores granulosa cell function and intercellular communication by regulating the IGF1R/Akt/mTOR signaling axis.

Dental-derived stromal cells (DSCs), including periodontal ligament stem cells, dental pulp stem cells, stem cells from the apical papilla, and stem cells from human exfoliated deciduous teeth, are promising candidates for oral and craniofacial regeneration because of their accessibility, expandability, and functional relevance to periodontal, dentin-pulp, bone, and neurovascular repair. However, the therapeutic performance of DSC-based products remains inconsistent, partly because conventionally expanded cells may be insufficiently adapted to the mechanical cues encountered in vivo. In this Review, we present mechanical preconditioning as a mechanomedicine-guided strategy for ex vivo functional priming of DSCs. We summarize how major DSC populations respond to defined biophysical cues such as tensile and compressive forces, fluid shear stress, hydrostatic pressure, matrix stiffness, and surface topography, and we discuss the principal mechanotransduction pathways involved. We further outline representative quantitative loading windows and consider how these may support subtype-specific and indication-specific preconditioning design. Finally, we highlight key translational barriers, including stromal cell heterogeneity, donor variability, senescence, uncertain persistence of mechanically induced states, safety concerns, and the lack of standardized manufacturing workflows. Overall, clinical translation will require a shift from descriptive mechanobiology toward parameter-defined, indication-specific, and good manufacturing practice-compatible preconditioning strategies for DSC-based regeneration.

Developing regenerative therapies to restore kidney function in patients with progressive renal disease represents a major challenge for modern molecular nephrology. Kidney organoids, three-dimensional kidney-like structures, which can now be generated by the directed differentiation of human pluripotent stem cells, have emerged as a powerful tool to study kidney development, physiology, and mechanisms of renal disease in vitro. Ultimately, kidney organoids may serve as an experimental platform to unravel the pathomechanisms of renal fibrosis and to test regenerative treatment approaches targeting fibrotic kidney diseases. However, the fibrotic phenotype in kidney organoids and its utility as a disease model remain to be fully characterized. Three-dimensional self-organizing kidney organoids containing nephrons and stromal cells were exposed to TGF-β1 cytokine to induce fibrotic remodeling. Organoids were analyzed by RNA sequencing and histology. Activation of TGF-β1 signaling in kidney organoids induced hallmarks of human kidney fibrosis, such as tubular atrophy, glomerulosclerosis, and interstitial fibrosis. RNA sequencing highlighted differential regulation of key pathways in kidney fibrosis: epithelial-to-mesenchymal transition, inflammation, metabolism, and JAK/STAT signaling. We identified candidate mediators of kidney fibrosis such as the JAK-STAT downstream target PIM1. Inhibition of PIM1 with the small molecule AZD1208 attenuated fibrosis development in the organoids. Kidney organoids are an amenable system for modeling kidney fibrosis and may guide therapeutic discovery.

Chronic radiation cystitis (CRC) is a disabling late adverse effect of pelvic irradiation, characterized by urothelial barrier loss, persistent inflammation, vascular alteration, and progressive fibrosis. No curative treatment exists. Mesenchymal stromal cells (MSCs) are investigated for their regenerative potential, however their long-term mechanisms of action in CRC remain incompletely elucidated. CRC was induced in rats by localized bladder irradiation (40Gy). Animals were randomized into unirradiated controls, irradiated controls, and irradiated MSC-treated groups. The treated group received three intravenous injections of adipose-derived MSCs at 4.5, 5.0, and 5.5 months post-irradiation, during the pre-chronic phase. Bladders were analyzed up to 18 months post-irradiation by immunostaining and RT-qPCR for urothelial differentiation (Upk1a, Upk1b, Upk2, Upk3a, Upk3b), epithelial proliferation (CK14⁺/PCNA⁺) and fibrosis (collagen I and III). Immune activation (CD45⁺ leukocytes, neutrophils, mast cells, Gata3, Th1, Th2) was analyzed by flow cytometry and vascular integrity by cystoscopy and Cspg4, VegfA expression. At 18 months post-irradiation, irradiated bladders showed a 90% loss of UPK3 staining and hyperplasia (urothelium thickening) greater than 300μm result of basal cell hyperproliferation, a ninefold increase compared to controls. MSC treatment reduced hyperplasia by 66% compared to irradiated rats, restored gene expression of Upk1a and Upk3a to 80-90% unirradiated control levels and decreased proliferative CK14⁺/PCNA⁺ progenitors by 93%). Immune response was attenuated, with a 49% reduction in Th2-associated cell infiltration and downregulation of Gata3 expression (p < 0.05) relative to irradiated rats. A decrease in mast cell degranulation (61%) was observed at neurovascular interfaces. Vascular architecture was preserved, with VegfA expression normalization and reduction of cystoscopic vascular lesions. Fibrotic remodeling was limited, with less accumulation of collagen 1 and a maintained physiological collagen 1/3 balance in the detrusor. MSC treatment had durable, multi-compartment effects in CRC. It limited basal hyperplasia, restored urothelial lineage differentiation, selectively modulated adaptive immune activation and mast cell activity, and constrained collagen 1-driven fibrosis. Vascular architecture was partially preserved, which together supported urothelial homeostasis in CRC. These findings provide strong preclinical evidence supporting MSCs as a targeted long-term regenerative strategy for CRC.

Progenitor cell epithelial to mesenchymal transition (EMT) could impact healing in the epidermal-dermal junction of the equine hoof stratum internum (SI). The hypothesis of this study was that K14 + CD105+ equine hoof SI progenitor cells assume a mesenchymal phenotype in the presence of inflammatory interleukins in vitro. K14 + CD105+ progenitor cell percentages isolated from proliferative scarred (fibrous) and healthy SI were quantified. Decellularized matrix was prepared from and K14 + CD105+ cells localized in healthy SI. Gene expression (CD44, CD105, E-cadherin, N-cadherin, β-catenin, K1, K10, p63, TGF-β1, -β2, -β3) and E- and N-cadherin+ cell percentages were determined in continuously cultured cells. Revitalized K14 + CD105+ cells were cultured in stromal medium with or without interleukins 2 and 6 (IL medium) or on decellularized SI matrix in stromal medium. After 21 days, cell gene expression (K1, K10, p63, β-catenin, TGF-β1, -β2, -β3), E-cadherin + and K14 + cell percentages, and medium TGF-β1 levels were measured. Most cells from healthy and fibrous SI were K14 + CD105+, and K14 and CD105 antigens were present on cells in situ and in vitro. K14 + CD105+ equine hoof SI progenitor cells maintained the immunophenotype over multiple cell passages in vitro. E-cadherin+ cell percentages were higher and K1 expression lower in K14 + CD105+ versus unsorted cells and more K14 + CD105+ cells were E- versus N-cadherin+. K14 + CD105+ cells cultured on matrix had the highest E-cadherin+ cell percentage and p63 and K10 gene expression. Cell TGF-β2 and β-catenin expression was highest with stromal medium, and cell TGFβ-1 and TGF β-3 expression was lowest with IL medium and on matrix, respectively. Cell TGF-β2, TGF-β3 and β-catenin expression was lower on matrix versus in stromal medium. Cells in IL medium had higher TGF-β3 expression than those on matrix. TGF- β1 levels were lower in IL versus stromal medium. The cell genetic and antigen profiles suggest that inflammatory interleukins drive mesenchymal differentiation while healthy matrix supports epidermal differentiation. K14 + CD105+ progenitor cells from the SI epidermal-dermal niche provide a platform to elucidate progenitor cell EMT capabilities and restore normal tissue healing. These findings have important implications for improving treatment strategies for injuries and disease of the epidermal-dermal junction.

Cancer stem cells (CSCs) represent a minor but highly adaptable subpopulation within tumors that drives long-term growth, metastasis, and therapy resistance. Their ability to survive and regenerate under metabolic and therapeutic stress relies on a unique integration of energy flexibility, redox balance, and proteostatic programs. While bulk tumor cells typically favor aerobic glycolysis and high protein turnover, CSCs often exhibit elevated mitochondrial activity, fatty acid oxidation, and selective suppression of proteasome function. These metabolic features support quiescence, stress tolerance, and self-renewal. Beyond energy production, metabolic intermediates such as acetyl-CoA, succinate, and lactate serve as epigenetic cofactors, linking nutrient availability to chromatin remodeling and transcriptional plasticity. Reactive oxygen species and antioxidant responses further tune this balance, shaping the transition between glycolytic and oxidative CSC states. These intrinsic programs are continuously influenced by the tumor microenvironment, where hypoxia, cytokine-driven signaling, and metabolic coupling with stromal and immune cells modulate CSC metabolism and reinforce stemness. Despite rapid progress, major conceptual and methodological gaps still limit our understanding of CSC metabolism and this review highlights these unresolved issues and further outline key contextual factors-including tumor-intrinsic, microenvironmental, systemic, and metastatic cues-that shape CSC metabolism and help explain the divergent observations reported across studies. Understanding this network will be essential for designing combinatorial therapies that target CSC metabolism while accounting for their heterogeneity and plasticity.

Primary glomerular disease, particularly IgA Nephropathy (IgAN), is one of the leading causes of kidney disease burden. IgAN is characterized by IgA deposition in the mesangial area; this is a typical pathological feature that leads to nephritis symptoms. Though the cause is unknown, the quadripartite attack theory is generally accepted. There are many reasons, but the biggest is an imbalance in the immune system. Recently, some studies have shown that mesenchymal stem cells (MSCs) can regulate IgA production and clearance by controlling B cells, T cells, and mucosal epithelial cells. Currently, the standard treatment for IgAN primarily relies on supportive care and immunosuppression, but these approaches are ineffective and have serious side effects. In contrast, stem cell therapy is a promising option and shows great promise in diagnosing and treating various kidney diseases. This review provides an update on recent advances in IgAN using stem cell therapy, including disease mechanisms, treatment pathways, clinical trials, case reports, safety concerns, current challenges, and prospects.

Immune thrombocytopenia (ITP) is a heterogeneous autoimmune disorder characterized by increased platelet destruction and impaired megakaryopoiesis within a dysregulated bone marrow niche. Conventional therapies often achieve only transient platelet recovery, failing to restore immune tolerance, thereby underscoring the need for mechanism-based therapeutic strategies. Mesenchymal stem cells (MSCs) have emerged as promising candidates due to their ability to modulate immune responses and repair the hematopoietic microenvironment. This review synthesizes current evidence regarding the biological properties, immunomodulatory mechanisms, and therapeutic applications of MSCs in ITP, emphasizing intrinsic abnormalities of patient-derived MSCs and the corrective potential of exogenous MSCs from distinct tissue sources. It further integrates emerging insights into MSC functional heterogeneity, optimization of culture conditions, priming strategies, and cellular engineering approaches that may enhance therapeutic efficacy and safety. By highlighting the interplay between immune tolerance restoration and bone marrow niche remodeling, this review provides a translational framework that links mechanistic understanding to the future clinical development of MSC-based therapies for ITP.

Metanephric mesenchymal cells (MMCs) hold therapeutic potential for acute kidney injury (AKI), but their efficacy is limited, and the mechanisms underlying their action remain unclear. This study aimed to investigate whether catalpol-pretreated MMCs (MMCs-cata) could enhance the efficacy of AKI treatment by regulating key signaling pathways. An AKI model was established in C57BL6 mice via intraperitoneal injection of cisplatin, and the therapeutic effects of MMCs-cata were compared with those of untreated MMCs. RNA-Seq was performed to analyze differentially expressed genes, Western blotting and ELISA were used to measure VEGF-A levels in MMC-cata and the supernatants. An in vitro model of cisplatin-induced renal tubular epithelial cell injury was developed to investigate the signaling pathways upstream and downstream of VEGF-A. The role of VEGF-A/VEGFR2 was confirmed through experiments involving gene silencing, neutralizing antibodies, and VEGFR2 blockade. Additionally, molecular docking simulations and Western blotting were performed to explore the effects of catalpol on the Wnt signaling pathway. MMCs-cata significantly improved renal function in AKI model mice by suppressing inflammation, oxidative stress, and necroptosis. RNA-Seq, Western blotting and ELISA revealed the activation of VEGF-related genes in MMCs-cata along with elevated intracellular and supernatant VEGF-A levels. In both the in vivo and in vitro models, silencing VEGF-A in MMCs-cata, neutralizing VEGF-A in the supernatant, or blocking VEGFR2 in tubular epithelial cells abolished the protective effects of MMCs-cata, while exogenous VEGF-A supplementation alone exerted protective effects. Mechanistic studies indicated that MMCs-cata activated the p38 pathway and suppressed the STAT3 pathway. Molecular docking and Western blotting confirmed that catalpol binds to Wnt3A, activating the canonical Wnt pathway to drive VEGF-A secretion. Catalpol pretreatment enhances the therapeutic efficacy of MMCs by activating the canonical Wnt pathway to promote VEGF-A secretion. VEGF-A interacts with VEGFR2 on renal tubular epithelial cells, likely through both p38 pathway activation and STAT3 pathway inhibition, thereby suppressing inflammation, oxidative stress, and necroptosis to alleviate AKI. This study provides novel insights into the integration of traditional Chinese medicine components with stem cell therapy for AKI management.

Circadian rhythms are endogenous, transcription-translation feedback loops that align cellular activities with the 24-h light-dark cycle. Stem-cell populations across tissues exhibit circadian oscillations that influence their self-renewal, proliferation, and differentiation. Key developmental pathways (Wnt/β-catenin, Notch, and Hedgehog) are increasingly recognized as both regulators and targets of circadian machinery. This review synthesizes current knowledge on the bidirectional crosstalk between circadian clock components and major stem-cell regulatory pathways, and evaluates how this interplay shapes tissue homeostasis, regenerative capacity, and therapeutic potential. Literature examining molecular interfaces between circadian clock genes and Wnt, Notch, and Hedgehog signaling was surveyed, with emphasis on transcriptional regulation, chromatin dynamics, post-translational control, and functional outcomes for stem-cell behavior and regeneration. Evidence indicates that core clock components modulate stem-cell pathways through direct transcriptional control, shared enhancer architecture, altered chromatin accessibility, and rhythmic protein modification. In turn, Wnt, Notch, and Hedgehog signals feed back onto clock genes, influencing circadian amplitude and phase within stem-cell niches. Perturbation of this reciprocal regulation disrupts tissue maintenance, diminishes regenerative responses, alters metabolic equilibrium, and may promote tumorigenesis. Circadian oscillators act as temporal gatekeepers of stem-cell function. Mapping the molecular interfaces between clock genes and developmental signaling pathways reveals new opportunities to refine regenerative therapies. Chronotherapeutic strategies, i.e. timing interventions to intrinsic circadian phases may enhance the efficacy, precision, and safety of stem-cell-based treatments.