Mesenchymal stem cells (MSC) represent the main stromal component of the bone marrow (BM) niche and are crucial to maintain hematopoietic tissue homeostasis. MSC exhibits extraordinary and multiple properties. In terms of expanding potential and differentiation capacity, Wharton jelly MSC (WJ-MSC), derived from the umbilical cord, was described as being greater and more performing than MSC from BM or other sources. WJ-MSC mimics the hematopoietic niche and supports hematopoietic stem cells (HSC) expansion ex vivo. This study aimed to evaluate the effects of human WJ-MSC cocultured with HSC in a B-cell differentiation protocol. Remarkably, results highlight WJ-MSC use as a preferable feeder layer to efficiently support HSC commitment toward the B-lineage. Over 11 days of HSC coculture with WJ-MSC, B-cell genes (E2A, RAG1, RAG2, etc.) expression patterns and B-cell markers (CD19, immunoglobulin chain, etc.) acquisition were evidenced. WJ-MSc were also able to unlock the B-lineage differentiation blockade of the acute lymphoblastic leukemia cell line Nalm16. This model might provide a new strategy to support ex vivo B-cell differentiation using the powerful properties of WJ-MSC. This study implements a new approach to improve understanding of B-leukemogenesis and B-cell acute lymphoblastic leukemia (B-ALL) pathophysiology.

The objective of the study was to compare the role of Cryopreserved human amniotic membrane (cHAM) for early pulp biomineralisation in comparison to Biodentine using the tooth culture model. This study was conducted in two stages. The first stage aimed to establish the tooth culture model and assess the viability of pulp cells at 28 and 50 days. In the second stage, pulp capping was performed in the tooth culture model using cHAM and Biodentine. A total of 20 freshly extracted teeth (for orthodontic reasons) were used for pulp capping with either cHAM or Biodentine. The samples were evaluated for biomineralisation at 14 and 28 days using micro-computed tomography (micro-CT) and histological analysis. Morphometric data from micro-CT scans were analysed using the Chi-square test. The viability of pulp cells in the tooth culture model was 66.1% at 28 days and 38.4% at 50 days. Pulp capping with cHAM resulted in a thicker and larger volume of mineralisation at 28 days (720 μm, 0.7 mm3) compared to Biodentine (630 μm, 0.46 mm3). Histological analysis revealed that the dentine formed in the HAM group was homogeneous and continuous, while the Biodentine group showed discontinuous foci of mineralisation in the newly formed hard tissue. Within the study's limitations, it is observed that cHAM induces enhanced early pulpal mineralisation compared to Biodentine.

Human embryonic stem cell (hESC) can be differentiated into definitive endoderm (DE) through multiple branching lineage choices. Although different DE differentiation methods have been established, there are still several limitations, such as the yield of heterogeneous cell populations containing undifferentiated or non-DE cells. Therefore, this study aimed to suppress the alternate fates at branch points and establish a robust and highly efficient differentiation protocol for hESC-derived hepatocytes (hESC-Heps). We developed a two-step DE induction protocol. First, hESCs were treated with a GSK-3α/β inhibitor and an mTOR inhibitor combined with TGF-β activation to generate an anterior primitive streak (progenitor to endoderm). Subsequently, a BMP inhibitor combined with TGF-β activation was used to abolish the mesoderm lineage. The resulting DE cells were further differentiated into hESC-Heps to evaluate their functionality. By regulating the branching lineage choices, we established an efficient two-step method that yielded up to 96% DE cells with minimal expression of pluripotency and mesodermal markers. Notably, this method reduced the dosage of Activin A, which makes it cost-effective for future applications. The derived hESC-Heps exhibited mature hepatocyte characteristics, including glycogen storage, indocyanine green uptake, and cytochrome P450 activity. Additionally, these cells demonstrated robust liver-specific functions such as sensitive innate immune responses and permissiveness to hepatitis B virus infection. In summary, we developed a novel and cost-effective method that achieves high-purity DE by precisely modulating cell fate decisions in the early stages. The derived hESC-Heps can serve as a model for further studies, such as host-virus interaction and hepatotoxicity testing.

Overcoming remyelination failure is one of the main targets in therapeutic strategies for multiple sclerosis. This process requires the differentiation of oligodendrocyte precursor cells (OPCs) to mature myelinating oligodendrocytes (OLs), a process known to be controlled by thyroid hormone, nuclear receptors, and sonic hedgehog (SHH). Retinoid X receptor gamma (RXRg) is one of the nuclear receptors acting as a positive regulator of remyelination, but little is known about its mechanisms of function. Using transcriptomic and pharmacological analysis of primary neural stem cell-derived OPCs, we show that RXRg is involved in the induction of the thyroid hormone-driven differentiation process and in refining it toward an oligodendrogenic cell fate. RXRg also emerged as an important negative modulator of SHH expression and signaling, as Shh and additional genes from this pathway were found to be strongly upregulated in Rxrg-/- OPCs. An inhibition of SHH signaling by cyclopamine or GANT61 entirely normalized the differentiation deficit of Rxrg-/- OPCs, but also myelination of newly generated Rxrg-/- OLs. Such data indicate a key role of SHH hyperactivity in the oligodendrogenesis block associated with the absence of RXRg. Importantly, hyperactivation of the SHH pathway by purmorphamine or SAG inhibited the oligodendrogenesis and myelination potential of wild-type OPCs, indicating that SHH hyperactivity can also be a sufficient factor to block OPC differentiation. These results point to RXRg as an important regulator of SHH pathway signaling and underline the need of an optimal, fine-tuning of SHH signaling to assure successful oligodendrogenesis.

Chronic non-healing wounds pose a significant clinical challenge, driven by dysregulation of the "inflammation-immune-microbiome" triad. Traditional "debridement-anti-infection-coverage" approaches fail to break the vicious cycle of dysbiosis and immune dysfunction. Leveraging structural diversity and bioactivity, natural polysaccharides provide a versatile platform for multi-targeted intervention. This review systematically explores the mechanisms through which polysaccharides modulate the wound immune microenvironment, restructure microbial communities, and facilitate barrier repair. This interaction enables precise regulation of macrophage polarization, particularly the promotion of the M2 phenotype, as well as neutrophil function and adaptive immunity, thereby alleviating chronic inflammation. Moreover, polysaccharides utilize a variety of mechanisms to impact the microbiome, including direct antimicrobial effects through electrostatic interactions and prebiotic support that promotes the colonization and metabolism of beneficial bacteria. This review also explores advancements in intelligent delivery systems, such as microenvironment-responsive hydrogels, discusses challenges in clinical translation, and considers future directions that incorporate single-cell multi-omics, microbiota-based personalization, organ-on-a-chip models, and phage-polysaccharide synergistic therapies. This work offers a theoretical foundation and translational perspective for the development of next-generation polysaccharide-based strategies for chronic wound management.

Polysaccharides have attracted increasing attention due to their potential protective effects against oxidative stress-related ovarian dysfunction. Panax ginseng (P. ginseng) stem-leaf contains abundant polysaccharides. However, their protective effects on ovarian granulosa cells remain underexplored. An acidic polysaccharide (PGPW-a2, 9535 Da) was isolated from P. ginseng stem-leaf. Its backbone consists of 4-α-GalAp, 2-α-Rhap, 2,4-α-Rhap, 3,4-α-Glcp, and 3,6-β-Galp residues, with three side chain (R1, R2 and R3). PGPW-a2 significantly boosted the functionality of antioxidant enzymes and reduced ROS levels by activating the Keap1/Nrf2/HO-1 signaling axis, and protected H2O2-damaged KGN cells by inhibiting the mitochondrial apoptosis pathway. Critically, these protective effects were substantially reversed by the Nrf2 inhibitor ML385. Furthermore, at equivalent concentrations, the main chain core unit of PGPW-a2 (PGPW-a2-a6) exhibited significantly attenuated antioxidant and anti-apoptotic effects compared to PGPW-a2, suggesting synergistic action between the main and branched chains is essential for full bioactivity. These findings indicate that PGPW-a2 protects KGN cells against oxidative stress, highlighting its potential relevance to ovarian health.

CAR T cell therapy has demonstrated remarkable efficacy in treating haematological malignancies, including B-cell lymphomas, B-cell leukaemias, and multiple myeloma. CAR T cell therapy for acute myeloid leukaemia (AML) is also urgently needed. One of the major challenges is identifying AML-specific antigens, since many potential candidates (e.g. CD33, CD123, CLL-1, CD70, TIM-3 and FLT3) are also expressed on normal haematopoietic progenitors. This can lead to 'on-target/off-tumour' toxicity and bone marrow aplasia. CAR NK cell therapy for AML shows promise as a lower-toxicity, off-the-shelf alternative. NK cells have a lower inherent risk of GVHD and may cause milder CRS/ICANS. In this review, we will describe the current status of CAR T/NK cell development for AML. We will also introduce a new CAR T-cell or NK-cell therapy that targets mismatched HLA-DRB1 in patients with AML who have relapsed following an allogeneic haematopoietic stem cell transplant.

The mechanistic role of trisomy 8 in the development of myelodysplastic syndrome (MDS) remains poorly defined. Here, we generated a trisomy 8 mouse model by transferring a human chromosome 8 into murine embryonic stem cells and prospectively examined the effects on hematopoietic stem cells (HSC) by trisomy 8. The expression of inflammatory genes was enhanced, and hematopoietic programs mediated by transcription factors and polycomb repressive complex 2 (PRC2) were dysregulated in trisomy 8 HSC, which impaired their self-renewal and balanced differentiation. Trisomy 8 HSC altered the chromatin accessibility and conformations and activated Y chromosome genes, such as Uty/Kdm6c epigenetic modifier, which is known to demethylate histone H3K27me3 modification. The Uty gene facilitated the activation of PRC2-target and Runx1-target genes in leukemogenesis and drove the proliferation of human trisomy 8 leukemic cells. Since the RUNX1 gene is frequently mutated in patients with trisomy 8 MDS, its deletion attenuated the enhanced expression of inflammatory genes and mitigated the impaired self-renewal of trisomy 8 HSC in mice. Our findings reveal that trisomy 8 altered the transcriptional programs and chromatin conformations in HSC and drove a pre-malignant state through activating the expression of Uty, suggesting a route for the development of trisomy 8 MDS.

Transendothelial migration (TEM), is a complex, multistep process impacted by diseases like autoimmune disorders and cancer. Deciphering aspects of the process enhances our understanding and possibilities for disease treatments. The extent to which this potential can be leveraged is often limited due to conventional assays neither accurately mimicking specific in vivo conditions like sheer stress nor visualizing the whole transmigration cascade, hence missing distinct mechanisms. Flow-based adhesion assays overcome these limitations and allow the use of various endothelial cells from different tissues with distinct properties. So far, a broader and translational assay application is hampered by potential operator-based bias/lack of standardization, as well as poor scalability due to time-consuming manual analysis. In this study, we successfully combined this assay with AI-based analysis including subsequent classification of cell-transmigration phases by a Keras/TensorFlow-based deep learning model. Trained on healthy-donor and pancreatic cancer patient-derived T cells, the model achieved a high accuracy of 91.6 % in identifying/categorizing cell transmigration, surpassing the currently accepted 80 %-threshold, therefore qualifying as a fast, standardized AI-based live-cell imaging tool. Additionally, its architecture grants highly convenient reconfiguration for various disease-model investigations. Hence, by combining an affordable and simplistic, yet potent live-cell-imaging technique with a comprehensive AI-approach, we have established a powerful tool which allows for integrating TEM-assays into various disease models.

CD34 has long been defined as a canonical marker for endothelial progenitors as well as hematopoietic stem cells, implicating its role in vascular development and hematopoiesis. However, the precise developmental hierarchy and lineage potential of CD34+ cells remain controversial. In this study, we integrated inducible genetic lineage tracing techniques, proteomics and single-cell RNA-seq (scRNA-seq) analyses to elucidate the dynamic developmental trajectory of CD34+ cells during various embryonic periods in both humans and mice. Remarkably, our analyses indicated that the progeny of CD34+ cells marked distinct, spatiotemporally restricted progenitor waves with divergent fates, at which point cells adopted endothelial, hematopoietic and fibroblastic fates, respectively. During gastrulation (E6.5-E8.5), an initial wave of CD34+ progenitors predominantly orchestrates vasculogenesis via a Kdr-dependent mechanism. Subsequently, from E9.5 to E14.5, cell cycle activation serves as a molecular switch, facilitating the endothelial-to-hematopoietic transition (EHT) of CD34+ progenitors. Unexpectedly, we identify a wave of CD34+ progenitors in late embryogenesis that gives rise to fibroblasts, distinct from earlier endothelial or hematopoietic lineages. Furthermore, because umbilical cord blood is a valuable source of different circulating stem/progenitor cells, we distinguish circulating endothelial progenitors from fibroblast progenitors in human cord blood by unique molecular signatures, with GFPT2 specifically marking the fibroblast progenitors. Collectively, our study provides a high-resolution spatiotemporal atlas of CD34+ cells during embryogenesis, redefining the temporal shifts of CD34+ cells in cell states and offering a precise framework for manipulating CD34+ cells in regenerative medicine.

Melanocyte stem cells (McSCs) are a crucial melanocyte reservoir within the hair follicle niche. This review provides an overview of the processes for McSC quiescence and activation. Because McSCs closely interact with hair follicle stem cells, we have focused on this interaction. Given the high prevalence of hair graying, the McSC system serves as a model for cellular aging. Here, we highlight current research on the mechanisms of hair graying.

Iron metabolism is increasingly recognized as a key player in the development and progression of various cancers. Iron is required for vital cellular processes such as energy production; however, it can also interact with reactive oxygen species to cause cellular toxicity. Consequently, a host of proteins coordinate iron homeostasis, and ferritin stands out as a promising therapeutic target due to its pivotal role in buffering cellular iron levels. This review explores the relevance of ferritin in brain cancers, shedding light on how it influences the biology of both tumor cells and cancer stem cells (CSCs), a population of tumor cells that is notable in their resistance to conventional treatment strategies. Ferritin plays a critical role in protecting against oxidative stress and boosting resistance to ferroptosis, a form of cell death often evaded by CSCs. Development of cutting-edge strategies designed to target ferritin, including ferritinophagy-inducing compounds and novel redox-based therapies that can capitalize on the iron dependency of CSCs is discussed in context. We propose that the iron addiction of brain cancer cells provides a specific susceptibility, whereby removing their iron buffering mechanism via targeting of ferritin can result in favorable treatment outcomes, including the induction of iron-dependent cell death. Future studies on the modulation of ferritin offer a ground-breaking therapeutic strategy to undermine CSC-driven tumor growth, overcome resistance to conventional therapies, and ultimately improve treatment outcomes for patients battling brain cancers.

Dental tissues development involves two distinct cell lineages: mesenchymal cells (derived from the cranial neural crest) and epithelial cells (derived from oral ectoderm and pharyngeal epithelium). Emerging evidence highlights the remarkable functional heterogeneity of cranial neural crest-derived dental mesenchymal stem cells (DMSCs), exhibiting pluripotency, self-renewal, and differentiation capacities. This heterogeneity enables a single DMSC population to generate specialized subpopulations with unique roles in teeth and periodontal tissues formation. Significant progress has been made in characterizing six major types of DMSCs and two populations of closely related cells: Tooth germ progenitor cells (TGPCs) and dental follicle stem cells (DFSCs), critical during early morphogenesis; Stem cells from human exfoliated deciduous teeth (SHEDs) and apical papilla stem cells (SCAPs), pivotal for root development; Dental pulp stem cells (DPSCs), periodontal ligament stem cells (PDLSCs), gingival mesenchymal stem cells (GMSCs) and alveolar bone mesenchymal stem cells (ABMSCs), essential for maintaining and regenerating mature dental tissues. A key breakthrough has unveiled the development and hierarchy of DMSCs by applying new techniques like single-cell RNA sequencing (scRNA-seq). To integrate insights into the development of teeth and periodontal tissues, this review synthesizes current knowledge on both developmental heterogeneity and subpopulation heterogeneity within DMSCs and related cells. These insights not only advance fundamental understanding of the developmental mechanisms of teeth and periodontal tissues, but also establish a promising framework for achieving more efficient tissue regeneration and repair engineering.

Diseases associated with obesity and metabolic dysregulation, such as diabetes and metabolic dysfunction-associated steatotic liver disease (MASLD) promote chronic low-grade inflammation, which in turn, may enhance the risk for cardiovascular disease. Emerging evidence in recent years suggests that chronicity of inflammation involves alterations in bone marrow homeostasis. Obesity-related inflammation and metabolic stress, including hyperglycemia or hyperlipidemia, may trigger rewiring of hematopoietic stem and progenitor cells (HSPCs) in the bone marrow, driving production of myeloid cells with heightened inflammatory capacity that in turn fuel and sustain chronic inflammation. This process is akin to trained immunity and may promote an inflammatory memory that links metabolic disorders to their cardiovascular complications. Clonal hematopoiesis of indeterminate potential (CHIP) is characterized by aging-related emergence of somatic mutations in hematopoietic cells that clonally expand and bear higher inflammatory potential. Importantly, a bidirectional link between CHIP and metabolic disorders as well as their cardiovascular sequelae emerges. Here, we review current concepts regarding the links between bone marrow biology and metabolic diseases and associated chronic inflammation.

Post-traumatic stress disorder (PTSD) is a debilitating neuropsychiatric condition triggered by severe trauma, characterised by dysregulated fear circuitry, hippocampal atrophy with impaired neurogenesis, chronic neuroinflammation, neuroendocrine dysregulation, and disrupted prefrontal-limbic connectivity. Existing treatments are largely symptomatic, failing to address underlying neurobiological deficits. Emerging regenerative approaches using human stem cells; particularly induced pluripotent stem cell-derived neural progenitor cells (iPSC-NPCs), human embryonic stem cells (hESCs), mesenchymal stem cells (MSCs), and their extracellular vesicles (EVs), offer mechanistic plausibility for repair through direct neuronal replacement, paracrine neurotrophic support (e.g., BDNF, GDNF, VEGF), immunomodulation (e.g., shifting microglia to anti-inflammatory phenotypes), and promotion of synaptic plasticity and epigenetic reprogramming. Preclinical evidence remains limited and largely indirect, with sparse PTSD-specific studies (e.g., one report of iPSC-NPC transplantation reducing fear behaviour and enhancing hippocampal BDNF/neuronal density in a rat model) supplemented by convergent data from adjacent CNS injury paradigms. MSC- and iPSC-derived EVs, enriched with regulatory miRNAs (e.g., miR-124, miR-21, miR-146a), emerge as a safer, cell-free alternative with strong immunomodulatory potential and greater translational feasibility. However, reproducibility is constrained by model variability, lack of independent replication, and absence of PTSD-focused clinical trials. Major challenges include tumorigenicity risks (especially for pluripotent-derived cells), immune rejection, epigenetic/genomic instability, manufacturing scalability, stringent regulatory requirements, and elevated ethical thresholds for invasive therapies in a non-lethal psychiatric disorder. This review examines how stem cell actions align with PTSD brain changes, critically assesses the limited evidence, and suggests a careful translational plan.

Nickel (Ni) is a naturally occurring heavy metal whose environmental levels have been steadily rising due to industrial activities and the widespread use of Ni-containing products. Ni exposure poses significant health risks, and studies in vertebrate models and human populations link Ni to developmental toxicity. However, the mechanisms by which Ni perturbs early developmental programs remain poorly understood. Here, we examined the effects of Ni exposure on pluripotency in mouse embryonic stem cells (mESCs) maintained under pluripotency-supporting conditions. Ni exposure led to aberrant upregulation of genes associated with mesodermal and endodermal lineages, while ectodermal gene expression remained largely unaffected. However, the expression of core pluripotency factors was preserved, indicating that Ni does not induce differentiation but instead disrupts normal transcriptional control within the pluripotent state. Mechanistically, Ni exposure caused a selective loss of the repressive histone modification H3K27me3 at bivalent promoters of mesoderm-associated genes without altering global H3K27me3 levels. Pharmacological inhibition of H3K27me3 demethylases attenuated Ni-induced gene activation, suggesting that localized H3K27me3 removal contributes to this aberrant activation of developmental genes. ESCs normally exist as heterogeneous populations that dynamically fluctuate between naïve and lineage-primed pluripotent states. Our findings indicate that Ni exposure perturbs this equilibrium through aberrant activation of lineage-associated genes while core pluripotency remains preserved. Such dysregulation of early transcriptional programs may predispose cells to abnormal fate decisions. These findings suggest a mechanistic link between Ni exposure and developmental abnormalities.

To explore the role of bone marrow mesenchymal stem cell-derived exosomes (BMSCs-Exos) in promoting the motility and functional enhancement of tendon-derived stem cells (TDSCs) and assess their potential in tendon regeneration and repair.

Multiple myeloma (MM) is a hematological malignancy that depends on the bone marrow microenvironment, and obesity, along with intramedullary adipocytes, is associated with an increased risk of developing MM. Adipocytes protect MM cells from chemotherapy by secreting adipokines and activating autophagy, while MM cells reprogram bone marrow adipocytes. Strategies to inhibit adipocyte lipolysis have been proposed as a novel approach for treating MM. MM progression is dependent on glutamine and glucose metabolism. Lipoproteins, particularly cholesterol levels, are becoming prognostic factors in MM. However, studies on lipids lack long-term paired data. Therefore, we analyzed long-term follow-up data from 115 autograft patients at Beijing Jishuitan Hospital over the past 7.5 years to investigate the long-term changes in lipid metabolism during the course of MM and the impact of efficacy. Teaser Abstract: Is blood cholesterol a gauge for myeloma relapse? A landmark study shows that with remission, cholesterol levels promptly rise, only to markedly drop upon the cancer's recurrence-unveiling a straightforward new approach to disease tracking.

With population aging, chemotherapy-induced thrombocytopenia (CIT) is a severe complication in elderly cancer patients, yet effective preventive and therapeutic strategies remain limited. Here, we demonstrate that dietary restriction (DR) significantly mitigates 5-fluorouracil (5-FU)-induced thrombocytopenia and promotes platelet recovery in both young and aged mice. Mechanistically, DR improves mitochondrial homeostasis in hematopoietic stem and progenitor cells and enhances their hematopoietic reconstitution capacity. This preconditioning facilitates mitochondrial activation after chemotherapy, thereby promoting megakaryocytic lineage recovery. Pharmacological mitochondrial activation in ad libitum-fed mice mimics the protective effects of DR, whereas mitochondrial inhibition in DR-treated mice markedly attenuates these benefits. Clinically, cancer patients with lower pre-chemotherapy body mass index ([BMI] 18.5-22.95 kg/m2) showed a lower incidence of CIT following 5-FU treatment than those with higher BMI. Together, we show that short-term DR significantly mitigates CIT and that targeting mitochondria may represent a novel therapeutic strategy for CIT in elderly cancer patients.