Embryonic macrophages emerge before the onset of definitive hematopoiesis, seed into discrete tissues and contribute to specialized resident macrophages throughout life. Presence of embryonic macrophages in the bone marrow and functional impact on hematopoietic stem cells (HSC) or the niche remains unclear. Here we show that bone marrow macrophages consist of two ontogenetically distinct cell populations from embryonic and adult origin. Newborn mice lacking embryonic macrophages have decreased HSC numbers in the bone marrow suggesting an important function for embryo-derived macrophages in orchestrating HSC trafficking around birth. The establishment of a normal cellular niche space in the bone marrow critically depends on embryonic macrophages that are important for the development of mesenchymal stromal cells, but not other non-hematopoietic niche cells, providing evidence for a specific role for embryo-derived macrophages in the establishment of the niche environment pivotal for the establishment of a normally sized HSC pool.

Fetal hematopoiesis takes place in the liver before colonizing the bone marrow where it will persist for life. This colonization is thought to be mediated by specification of a microenvironment that selectively recruits hematopoietic cells to the nascent bone marrow. The identity and mechanisms regulating the specification of this colonization niche are unclear. Here we identify a VCAM1+ sinusoidal colonization niche in the diaphysis that regulates neutrophil and hematopoietic stem cell colonization of the bone marrow. Using confocal microscopy, we find that colonizing hematopoietic stem and progenitor cells (HSPC) and myeloid cells selectively localize to a subset of VCAM1+ sinusoids in the center of the diaphysis. Vcam1 deletion in endothelial cells impairs hematopoietic colonization while depletion of yolk-sac-derived osteoclasts disrupts VCAM1+ expression, and impairs neutrophil and HSPC colonization to the bone marrow. Depletion of yolk-sac-derived myeloid cells increases fetal liver hematopoietic stem cell numbers, function and erythropoiesis independent of osteoclast activity. Thus, the yolk sac produces myeloid cells that have opposite roles in fetal hematopoiesis: while yolk-sac derived myeloid cells in the bone marrow promote hematopoietic colonization by specifying a VCAM1+ colonization niche, a different subset of yolk-sac-derived myeloid cells inhibits HSC in the fetal liver.

Mitochondria play a vital role in cellular energy metabolism and vascular health, with their function directly influencing endothelial cell (EC) bioenergetics and integrity. Mitochondrial transfer has emerged as a key mechanism of intercellular communication, impacting angiogenesis, tissue repair, and cellular homeostasis. This review highlights recent findings on mitochondrial transfer, including natural mechanisms - such as tunneling nanotubes (TNTs) and extracellular vesicles (EVs) - and artificial approaches like mitochondrial transplantation. These processes enhance EC function and support vascularization under pathological conditions, including ischemia. While early clinical trials demonstrate therapeutic potential, challenges such as mitochondrial instability and scaling host-derived mitochondria persist. Continued research is essential to optimize mitochondrial transfer and advance its application as a therapeutic strategy for restoring vascular health.

Nanodiamonds (NDs) display several attractive features rendering them useful for medical applications such as drug delivery. However, the interactions between NDs and the immune system remain poorly understood. Here, we investigated amino-, carboxyl-, and poly(ethylene glycol) (PEG)-terminated NDs with respect to primary human immune cells. We applied cytometry by time-of-flight (CyToF) to assess the impact on peripheral blood mononuclear cells at the single-cell level, and observed an expansion of plasmacytoid dendritic cells (pDCs) which are critically involved in antiviral responses. Subsequent experiments demonstrated that the NDs were actively internalized, leading to a vigorous type I interferon response involving endosomal Toll-like receptors. ND-NH2 and ND-COOH were more potent than ND-PEG, as evidenced by using TLR reporter cell lines. Computational studies demonstrated that NDs interacted with the ligand-binding domains of TLR7 and TLR9 with high affinity though this was less pronounced for ND-PEG. NDs with varying surface functionalities were also readily taken up by macrophages. To gain further insight, we performed RNA sequencing of a monocyte-like cell line exposed to NDs, and found that the phagosome maturation pathway was significantly affected. Indeed, evidence for lysosomal hyperacidification was obtained in dendritic cells and macrophages exposed to NDs. Moreover, using a reporter cell line, NDs were found to impinge on autophagic flux. However, NDs did not affect viability of any of the cell types studied. This study has shown that NDs subvert dendritic cells leading to an antiviral-like immune response. This has implications not only for drug delivery but also for anticancer vaccines using NDs.

Atrial fibrillation (AF) is the most common sustained arrhythmia. Clonal hematopoiesis of indeterminate potential (CHIP), characterized by the clonal expansion of hematopoietic stem cells due to acquired mutations without hematologic malignancies, has emerged as a potential risk factor for AF. This narrative review summarizes the shared risk factors between CHIP and AF, including age, lifestyle behaviors and cardiometabolic conditions. It then explores the underlying mechanisms including inflammation, atrial fibrosis and abnormal red cell distribution width. Among these, inflammation is a central driver that promotes abnormal calcium handling, which further accelerates atrial remodeling. For specific mutations, TET2 mutations correlate strongest with AF, with other mutations in genes such as ASXL1, JAK2, TP53, PPM1D and spliceosomes, may also modulate AF susceptibility, though their precise roles require further investigation.

Plants belonging to the Meliaceae family, such as Trichilia silvatica C. DC., known as catiguá-branco, have attracted considerable interest in phytochemical research due to their diverse and significant secondary metabolites. Trichilia silvatica has traditionally been employed in Brazilian medicine to treat inflammatory disorders. Moreover, studies have reported its antioxidant and antimicrobial properties, highlighting its potential therapeutic applications.

Chronic exposure to arsenic, a toxic metalloid frequently found in groundwater and food, represents a significant environmental health risk and has been implicated in the etiology of several cancers, including ovarian cancer. However, the precise pathways through which arsenic exerts its toxic impact on the ovary are not fully understood. This study investigates the impact of chronic arsenic exposure at environmentally relevant concentrations (75 ppb or μg/L) on primary human ovarian surface (OCE1) and fallopian tube (FNE1) cultures derived from the same donor. These heterogeneous cultures provide a unique, human-relevant platform to investigate how chronic arsenic exposure influences distinct cell types within a shared microenvironment. Prolonged arsenic exposure induced significant cytotoxicity and promoted the formation of giant and/or multinucleated cells in both cultures. These cells exhibited phagocytosis-like properties, actively engulfing apoptotic debris. Transcriptomic analyses and pathway enrichment revealed robust activation of pro-inflammatory signaling, notably the canonical NF-κB pathway. This was marked by nuclear translocation of the NF-κB p65 subunit and elevated expression and secretion of pro-inflammatory cytokines, including TNFα, IL-6, and IL-8, driving a sustained inflammatory response. Moreover, arsenic-exposed cells displayed persistent DNA damage, as indicated by increased γ-H2AX foci, accompanied by nuclear structural alterations and elevated expression of cancer stem cell markers, including OCT2, CD133, and ALDH1. These findings suggest that arsenic-induced inflammation and genomic instability converge to promote a tumor-supportive microenvironment, highlighting the potential role of chronic arsenic exposure in ovarian carcinogenesis, particularly in the context of inflammation-driven carcinogenesis.

Three-dimensional patient-derived organoids (PDOs) have emerged as a powerful model for investigating the molecular and cellular mechanisms underlying bladder cancer, particularly in the context of cancer stem cells (CSCs) and drug screening. However, a significant limitation of conventional PDOs is the absence of tumor microenvironment (TME), which includes critical stromal, immune and microbial components that influence tumor behavior and treatment response. In this review, we provide a comprehensive overview of the recent advancements in PDO co-culture systems designed to integrate TME elements. Additionally, we emphasize the role of biomedical engineering technologies, such as 3D bioprinting and organoids-on-a-chip, in enhancing the physiological relevance of these models. Furthermore, we explore how bladder PDO co-culture systems are applied in research on bladder CSC characterization, evolution and treatment responses. Finally, we discuss future directions for improving PDO systems to achieve more accurate preclinical modeling and drug discovery.

We previously identified tetraspanin 7 (Tspan7) as a candidate gene influencing body weight in an obesity-related gene screening study. However, the mechanisms underlying its involvement in body weight regulation remained unclear. This study aims to investigate the role of TSPAN7 from a metabolic perspective.

Esophageal cancer, a highly malignant tumor with poor prognosis, is characterized by the presence of cancer stem cells (CSCs) that drive tumor initiation, metastasis, and recurrence. This study investigates the molecular mechanism by which glucose transporter 1 (GLUT1) maintains esophageal CSC-like properties through regulation of autophagy-dependent ferroptosis via epidermal growth factor receptor (EGFR). Using shRNA to knock down GLUT1 or EGFR and constructing a GLUT1 overexpression vector in KYSE520 cells, we employed western blotting, qRT-PCR, flow cytometry, sphere formation, Transwell assays, and xenograft models to assess stemness markers (NANOG, OCT4, SOX2), autophagic flux (LC3B, P62, Beclin1), and ferroptosis-related parameters (ROS, Fe2+, GSH, GPX4, COX2). Mechanistic analyses included co-immunoprecipitation to validate the GLUT1-EGFR interaction, chloroquine to inhibit autophagy, and cycloheximide/MG132 to evaluate EGFR protein stability. Results showed that GLUT1 depletion reduced CSC marker expression, increased ROS and Fe2+ levels, depleted GSH, and induced lipid peroxidation, sensitizing cells to ferroptosis while activating autophagy (elevated LC3 II/I, Beclin1; reduced P62); autophagy inhibition exacerbated cell death, indicating a protective role for autophagy in this context. GLUT1 directly bound to EGFR, stabilizing the receptor by blocking ubiquitin-proteasome-mediated degradation, whereas EGFR knockdown enhanced autophagic flux and reversed GLUT1-overexpression-induced ferroptosis resistance and stemness maintenance. In vivo, GLUT1 knockdown suppressed tumor growth and lung metastasis, and clinical samples revealed a positive correlation between GLUT1 and EGFR expression, linked to advanced TNM stages and poor survival. Collectively, these findings demonstrate that GLUT1 preserves esophageal CSC-like characteristics by stabilizing EGFR to inhibit autophagy-dependent ferroptosis, highlighting targeting GLUT1 as a potential therapeutic strategy to eliminate CSCs and combat esophageal cancer progression.

We report the results of two independent, concurrently performed studies evaluating the safety and efficacy of donor-derived mesenchymal stromal cell (MSC) infusions in inducing immune-tolerance in nonhuman primate (NHP) and human kidney transplant recipients treated with depletional induction and belatacept/sirolimus maintenance. Fifteen NHPs received rhesus ATG induction and were divided in three groups: control (n=6), pre-transplant thymic irradiation (TI, n=4), and TI with monthly donor-MSC infusion (n=5). Sirolimus was discontinued at day-180, and belatacept at day-365 post-transplantation. In humans, six patients enrolled in ITN062ST underwent transplantation with alemtuzumab induction; four received 12 monthly donor-MSC infusions followed by immunosuppression withdrawal (ISW) if eligible. Donor-MSC infusion was acutely well-tolerated in humans and NHPs. Chimerism was not established, and tolerance was not induced in either study. Two of five NHPs that received MSCs rejected while on belatacept monotherapy with detectable donor-specific antibody (DSA). Two patients did not initiate ISW due to de novo DSA and borderline rejection, and two patients failed ISW due to reversible rejection. In conclusion, donor-MSCs can be given to NHPs or humans repeatedly without acute consequences, but they neither lead to detectable chimerism nor induce tolerance. In a subset of recipients, infused MSCs can be sensitizing. Trial Registration. ClinicalTrials.gov - NCT03504241.

Paeonia lactiflora is an important garden ornamental flower and has become a new cut flower in the international market in recent years. Stem mechanical strength is an important index of cut flower quality. P. lactiflora cultivars possess varying stem mechanical strength, which directly result in different cut P. lactiflora quality. In our previous study, a significant difference in phloem callose deposition was detected between two P. lactiflora cultivars with varying stem mechanical strength. However, the relationship and interaction mechanism between the phloem callose and mechanical strength in P. lactiflora stems are still unclear. This study aimed to analyze the effects of reduced phloem callose on the mechanical strength in P. lactiflora stems. The results revealed a negative correlation between the callose content and mechanical strength in P. lactiflora stems during early developmental stages. Exogenous 2DDG treatment led to a 13.6% reduction in the callose content, concomitantly enhancing the stem mechanical strength by 14.2% via facilitating the formation and secondary cell wall (SCW) thickening of xylem fiber cells. Further investigations showed that exogenous 2DDG treatment promoted carbon transport in P. lactiflora stems, thereby increasing the carbon content. Additionally, it induced alterations in the metabolism of lipids, amino acids, and organic acids in the stems and spurred the diffusion of lipids from the pith to the xylem. Consequently, reducing the content of callose improved the formation and SCW thickening of xylem fiber cells by modifying material transport and metabolism, thereby enhancing the stem mechanical strength of P. lactiflora. This study offers novel perspectives regarding the regulatory mechanism of P. lactiflora stem mechanical strength.

Human induced pluripotent stem cells (iPSCs) can be differentiated into hepatocyte-like cells (iPS-Heps); however, maintaining the long-term proliferation and hepatic characteristics of iPS-Heps remains a challenge. In this study, we aimed to develop a human iPSC-derived hepatic organoid (iHO) culture system that effectively retains hepatic characteristics long term. Our original culture strategy, using bile acids and their receptor (farnesoid X receptor [FXR]) agonists, yielded human iHOs capable of long-term culture with a distinctive "grape-like" structure. Comprehensive analysis showed that these iHOs maintained hepatocyte-like phenotypes, even after multiple passages, whose gene expression profiles were consistent with those of fetal hepatocytes. In addition, the overexpression of small heterodimer partner (SHP), a downstream gene of FXR, in iHOs negatively regulated genes related to the intestine and cholangiocytes. Our data demonstrated that bile acid-FXR signaling promotes both the hepatic characteristics and proliferative potential of iHOs, offering promising potential for future applications in regenerative medicine and as a disease model.

There has been significant progress in the clinical development of allogeneic off-the-shelf chimeric antigen receptor (CAR)-engineered cell therapies for the treatment of cancer and autoimmune diseases. Unlike autologous CAR cell therapies, allogeneic approaches overcome challenges such as high costs, labor-intensive manufacturing, and stringent patient selection. This makes allogeneic therapies a more universally applicable option for a diverse patient population. In this review, we examine recent clinical advancements in allogeneic CAR cell therapies, including CAR-T cell therapy derived from healthy donor peripheral blood mononuclear cells, as well as CAR-NK cell therapy from cord blood or induced pluripotent stem cells. We provide an overview of their genetic engineering strategies, clinical designs, and outcomes, highlighting their promising efficacy and safety. Additionally, we summarize key preclinical developments, address key challenges, and explore future directions to provide insights into emerging trends in the field.

Chronic pain is a global health issue, yet effective treatments remain limited due to poor preclinical-to-human translation. To address this, we developed a high-content screening (HCS) platform using hiPSC-derived nociceptors to identify analgesics targeting the peripheral nervous system. These cells, cultured on multi-well microelectrode arrays, achieved nearly 100% active electrodes by week 2, maintaining stable activity for at least 2 weeks. After 28 days, we assessed drug effects on neuronal activity, achieving strong assay performance (robust Z' > 0.5). Pharmacological tests confirmed responses to key analgesic targets, including ion channels (Nav, Cav, Kv, and TRPV1), neurotransmitter receptors (AMPAR and GABA-R), and kinase inhibitors (tyrosine and JAK1/2). Transcriptomic analysis validated target expression, though levels differed from primary human DRG cells. The platform was used to screen over 700 natural compounds, demonstrating its potential for analgesic discovery. This HCS platform facilitates the rapid discovery of uncharacterized analgesics, reducing preclinical-to-human translation failure.

The attempts are being made to investigate the new approaches to identify and treat the chemical-induced neurotoxicity. The human mesenchymal stem cell (hMSC) secretome has been recognized as one of the promising approaches, as it is rich in bioactive factors that promote regeneration and neuroprotection. We examined the neuroprotective effects of stimulated and unstimulated hMSC-secretomes on human iPSC-derived neural progenitor cells (hNPCs) exposed to pesticide-monocrotophos (MCP). In-vitro assays were employed to assess the neuroprotective potential of MSC secretomes on hNPCs exposed to subtoxic concentrations of MCP. Comprehensive multi-omics analyses (proteomics and transcriptomics), bioenergetics assessments, and computational bioinformatics analyses were performed to elucidate the underlying molecular mechanisms and therapeutic effects. As anticipated, MCP exposure decreased viability, caused morphological changes, increased oxidative stress, and disrupted mitochondrial function in hNPCs. The treatment with MSC secretomes at 50 % concentration restored cell viability, morphology, and oxidative stress markers to near-normal levels. Bioenergetics analyses revealed significant improvements in mitochondrial oxygen consumption rates, ATP production, and spare respiratory capacity following secretome treatment, which was corroborated by proteomic analyses indicating restoration of mitochondrial protein expression and function. Transcriptomic profiling identified critical MCP-dysregulated miRNAs (including hsa-miR-138-5p and hsa-miR-219a-5p) and their inverse relationship with altered protein expression levels, highlighting the regulatory capacity of hMSC secretomes. The study demonstrates the therapeutic potential of MSC secretomes in mitigating chemical-induced developmental neurotoxicity by modulating oxidative stress, mitochondrial recovery, and miRNA-mediated signaling. Stimulated hMSC secretomes, which are enriched with bioactive molecules, showed enhanced efficacy, making them promising candidates for targeted therapies in chemical neurotoxicity interventions.

Regenerative medicine is a new hope for the treatment of diseases such as polycystic ovarian syndrome (PCOS). Due to the beneficial effects of adipose stem cell-derived conditioned media transplantation (ASC-CM) on polycystic ovary phenotypes and also considering the relationship between uterine function and ovarian hormone level, this study, for the first time, evaluates the transplantation of ASC-CM in compared to ASC on the altered state of the uterus, estrogen level as well as its receptors expression in PCOS model. Animals were divided randomly into two groups, including the CMC group and the PCOS group. After PCOS confirmation, the PCOS rats were allocated into four subgroups, including PCOS-Vehicle, PCOS-Media, PCOS-ASC, and PCOS-ASC-CM. After four weeks, the uterus tissues were removed for weighing, histomorphometrical evaluation, and immunoblotting assessments of estrogen receptors (ERα and ERβ). The sera were obtained for estrogen hormone level evaluation. Uterine histopathological conditions, such as marked reductions of the heights of the luminal and glandular epithelial cells as well as altered lining epithelial cells of the lumen and glands, alongside diminished uterine weight and thickness, were observed in the PCOS-V and PCOS-Media groups when compared to those of the CMC group. Additionally, their estrogen levels and ERα and ERβ expression levels decreased. However, ASC-CM administration, compared to ASCs alone, more effectively improved these altered uterine conditions of PCOS rats, resulting in enhanced uterine and endometrial wall thickness, luminal and glandular epithelial cell heights, estrogen levels, and also ESRα and ESRβ mRNA transcripts expression compared to those of the PCOS-V and PCOS-Media groups. So, the transplantation of ASC-CM, compared to ASC, could induce more estrogenic histological changes in the PCOS uterine alterations, which suggests a helpful strategy for improving forgotten uterine status in PCOS phenotypes.

Bone marrow mesenchymal stem cells (BMSCs) are stem cells that reside in bone marrow and have multidirectional differentiation potential. BMSCs have been used to treat bone injury. However, long-term passage leads to the aging of BMSCs and the weakening of osteogenic differentiation. Furthermore, brain-derived neurotrophic factor (BDNF) may enhance the antiaging ability of BMSCs. The purpose of this study was to investigate the role of BDNF in the senescence and osteogenic differentiation of human BMSCs (hBMSCs).