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).

iPSC-based models are valuable for studying the mechanisms and potential treatments of mitochondrial disorders. We generated two iPSC lines from fibroblasts of a patient with a novel MT-ATP6/8 mutation (m.8570 T > C). The infant was diagnosed with a mitochondrial disease featuring cardiac hypertrophy, brain atrophy, developmental delay, and metabolic crises with elevated lactate. Mutation heteroplasmy in blood leukocytes was 95 %. Leigh syndrome-like cranial MRI abnormalities were absent at 4 months of age. We introduced reprogramming factors by Sendai virus and assessed the pluripotency of the resulting iPSCs. As control iPSC-line, we characterized the CRMi004-A line from the RUCDR repository.

To investigate whether bone marrow mesenchymal stem cells derived exosomes (BMSCs-exo) can alleviate sepsis-induced lung injury and its related mechanism by inhibiting ferroptosis of macrophages.

The derivation of porcine embryonic stem cell (pESC) lines remains a major challenge in this field. To date, the porcine naïve ESCs have yet to be successfully established, and standardized criteria for their characterization and evaluation are still lacking. The regulation of X-chromosome activity integrates information from embryonic development and the dosage of sex chromosomes, which is closely associated with the pluripotent state of embryonic stem cells. In this study, we aimed to establish pESC lines in LCDM medium from porcine blastocyst-stage embryos, and analyzed the features of ESCs from the sight of X chromosome activity. We assessed molecular markers and epigenetic characteristics to confirm pluripotency and X chromosome activity in porcine parthenogenetic ESCs (named as ppLCDM) using XIST RNA-FISH, immunofluorescence staining, single-cell RNA sequencing (scRNA-seq), and other techniques. Results showed that ppLCDM cells expressed most pluripotent markers. The percentage of ppLCDM cells exhibiting H3K27me3 and XIST aggregation signals increased with passage, indicating the progressive establishment of X-chromosome inactivation (XCI). Meanwhile, the pluripotency of most ppLCDM cells gradually declined during extended passaging. However, two distinct patterns of ppLCDM cells were observed from passage 35 (type I cells, P35-I) displayed normal XCI states, while type II cells (P35-II) exhibited X-chromosome erosion-like state, characterized by the loss of aggregation signals, abnormal X-linked gene ratios. Particularly, the pluripotency of ppLCDM cells with an X-chromosome erosion-like state undergoes unusual changes compared to normal cells. These findings indicate that X chromosome activity is closely associated with the pluripotent state of porcine ESCs and that heterogeneity in X chromosome activity arises during passaging. Our research provides crucial insights into X chromosome dynamics in large-animal ESC models and contribute to ongoing efforts to establish stable naïve pESC lines.

Recent work has shown that prolonged expression of recombinant proteins after adeno-associated virus (AAV)-mediated delivery of gene therapy to long-lived, ventricle-lining ependymal cells can profoundly affect disease phenotypes in animal models of neurodegenerative diseases. Here, we performed in vivo screens of millions of peptide-modified capsid variants of AAV1, AAV2, and AAV9 parental serotypes in adult nonhuman primates (NHPs) to identify capsids with potent transduction of key brain tissues, including ependyma, after intracerebroventricular injection. Through these screens, we identified an AAV capsid, AAV-Ep+, with markedly increased potency in transducing ependymal cells and cerebral neurons in NHPs. AAV-Ep+'s potency was conserved in three species of NHP, two mouse strains, and human neurons derived from induced pluripotent stem cells. To apply AAV-Ep+ to the treatment of ceroid lipofuscinosis type 2 disease, a lysosomal storage disorder caused by loss-of-function mutations in tripeptidyl-peptidase 1 (TPP1), we used the capsid to package the human TPP1 transgene (AAV-Ep+.hTPP1) and delivered the construct by intracerebroventricular injection into mice lacking TPP1 activity. AAV-Ep+ provided robust and therapeutically relevant TPP1 protein concentrations in these mice, significantly improving tremor and life span. In NHPs, high cerebrospinal fluid (CSF) TPP1 concentrations were achieved after intracerebroventricular delivery of AAV-Ep+.hTPP1 at a total dose of 1 × 1012 viral genomes, which was more than 30× lower than previously reported doses in NHPs. These results suggest that AAV-Ep+ may be a potent vector for gene therapy applications where CSF protein expression is required.

During asymmetric cell division (ACD) of radial glia progenitors (RGPs), the cortical polarity regulator Par-3 is detected in the cytoplasm colocalizing with dynein and Notch ligand DeltaD (Dld). What drives Par-3 to the cytoplasm and its impact on RGP ACD remain unknown. Here, we visualize cytoplasmic Par-3 using in vivo time-lapse imaging and find that Ser954 of zebrafish Par-3 is phosphorylated by Aurora kinase A (AurkA) in vitro. Expression of the nonphosphorylated mutant Par-3S954A dominant negatively affects embryonic development, reduces cytoplasmic Par-3, and disrupts the anteroposterior asymmetry of cortical Par-3 and Dld endosomes and, in turn, daughter cell fate. AurkA in mitotic RGPs shows dynamic pericentrosomal distribution that transiently colocalizes with cortical Par-3 preferentially on the posterior side. AurkA is both necessary and sufficient to increase cytoplasmic while decreasing cortical Par-3, disrupts Par-3 cortical asymmetry, and perturbs polarized Dld endosome dynamics. These findings suggest that AurkA regulates Par-3 cortical-cytoplasmic dynamics that is critical for ACD and daughter cell fate.

The newly generated CD4 single-positive (SP) T lymphocytes are featured by enhanced IL-4 but repressed IFN-γ production. The mechanisms underlying this functional bias remain elusive. Previous studies have reported that CD4+ T cells from mice harboring dendritic cell (DC)-specific deletion of IL-27p28 display an increased capacity of IFN-γ production upon TCR stimulation. Here, we demonstrated that similarly altered functionality occurred in CD4SP thymocytes, recent thymic emigrants (RTEs), as well as naive T cells from either Cd11c-p28f/f mice or mice deficient in the α subunit of IL-27 receptor. Therefore, DC-derived IL-27p28-triggered, IL-27Rα-mediated signal is critically involved in the establishment of functional bias against IFN-γ production during their development in the thymus. Epigenetic analyses indicated reduced DNA methylation of the Ifng locus and increased trimethylation of H3K4 at both Ifng and Tbx21 loci in CD4SP thymocytes from Cd11c-p28f/f mice. Transcriptome profiling demonstrated that Il27p28 ablation resulted in the coordinated up-regulation of STAT1-activated genes. Concurrently, STAT1 was found to be constitutively activated. Moreover, we observed increased accumulation of STAT1 at the Ifng and Tbx21 loci and a strong correlation between STAT1 binding and H3K4me3 modification of these loci. Of note, Il27p28 deficiency exacerbated the autoimmune phenotype of Aire-/- mice. Collectively, this study reveals a novel mechanism underlying the functional bias of newly generated CD4+ T cells and the potential relevance of such a bias in autoimmunity.

Amniotic membrane transplantation (AMT) has emerged as a versatile therapeutic modality with significant applications in wound healing, tissue regeneration, and ophthalmology. This review comprehensively evaluates AMT's efficacy in acute and chronic wound management, where it has been shown to alleviate pain, reduce infection risk, and facilitate epithelialization. In chronic wounds, AMT enhances healing through mechanisms such as re-epithelialization, angiogenesis, and immune modulation. Additionally, AMT exhibits promise in nerve regeneration, demonstrating potential in the repair of peripheral and central nervous system injuries by fostering neural recovery and minimizing scar formation. In ophthalmology, AMT is instrumental in corneal surface reconstruction, conjunctival repairs, and the management of dry eye syndrome and limbal stem cell deficiency. While the benefits of AMT are well-documented, this review also addresses significant challenges, including variability in success rates across different clinical conditions, ethical concerns regarding donor tissue usage, and regulatory hurdles impacting its broader clinical application. Furthermore, we integrate recent advances in the understanding of AMT's molecular mechanisms - such as its antioxidant effects via Nrf2/HO-1 pathway and immune modulation via P2X7 receptor pathways - and highlight innovative strategies including the incorporation of nanoceria nanoparticles, Vitamin D3 supplementation, and gene therapy approaches to enhance AMT outcomes. By exploring these dimensions, the review highlights not only the current state of AMT but also its potential future role in advancing regenerative medicine, including emerging applications in spinal cord repair, orthopaedics, and tissue engineering. This updated synthesis aims to inform clinicians and researchers about the multifaceted applications of AMT, promoting further investigation and optimization of this promising therapeutic approach.

A primary therapeutic characteristic of mesenchymal stem/stromal cells (MSCs) is their immunomodulatory activity. Adipose-derived stem/stromal cells (ASCs) are an abundant and easily isolated source of MSCs shown to have high immunosuppressive activity, making them attractive for therapy. Understanding the heterogeneous immunomodulatory potential of ASCs within the stromal vascular fraction (SVF) of adipose tissue could better inform treatment strategies. In this study, we integrate single-cell RNA sequencing (scRNA seq) with bulk proteomics to characterize subpopulations of SVF-derived ASCs that are phenotypically similar to cytokine-licensed, cultured ASCs. To better define the licensing process, we present scRNA seq and bulk proteomics data of cultured (P2) ASCs exposed to inflammatory cytokines, showing enrichment of pathways related to inflammation and apoptosis that positively correlate to the cytokine-mediated, trajectory-derived pseudotime. Using the Scissor algorithm, we integrate the proteomics data with uncultured (P0) SVF scRNA seq data, identifying an ASC subpopulation that is phenotypically like the cytokine-stimulated ASCs (Scissor-positive). Interactome analysis identifies Scissor-positive ASCs as stress adaptive immune regulators that function through IL6 and broad SEMA4 interactions and higher Visfatin signaling, while Scissor-negative ASCs show strong signatures of ECM remodeling through FN1 and immunosuppression through THY1 and MIF signaling. Our multimodal, integrative approach enabled identification of previously unidentified, distinct ASC subpopulations with differing immunomodulatory phenotypes that are present in, and can potentially be selected from, P0 SVF ASCs.

Mesenchymal Stem cells (MSCs) have a wide range of therapeutic applications due to their self-renewal and multi-lineage differentiation ability; large-scale production of MSCs is possible only with a highly efficient medium, which facilitates increased proliferation of MSCs within a short period. Recently, Human Platelet Lysate (hPL) has emerged as a promising substitute for fetal bovine serum (FBS) for cell expansion. The goal of this study is to optimize a stable gel formulation for the 3D expansion of MSCs using hPL as a matrix material for the improved proliferation of Human Umbilical Cord Tissue derived MSCs (hUCT-MSCs) in comparison to FBS and hPL-supplemented media in 2D culture. To assess the potential benefits of the hPL gel system, in promoting cell proliferation capacity, hUCT-MSCs were cultured on hPL gel coated-dish supplemented with hPL CM, and in FBS CM. Among the varying concentrations, 20% hPL gel was optimized to have more functional stability and shorter gelation time. SEM analysis and gel degradation study at different concentrations revealed the structural integrity and morphology of the gel. Microscopic images and histological staining by H&E were conducted to understand the multi-layered proliferation of hUCT-MSCs in hPL Gel. Flow cytometry analysis reported the expression of positive markers for human umbilical cord MSCs, namely CD 90+ and CD 105+, in hPL Gel and hPL Complete Medium (CM) similar to that in FBS. The CCK8 Assay carried out for each culture system, generated OD values respective to cell viability and proliferation. OD values of 1.65 nm, 1.27 nm, and 0.92 nm on average were observed for hPL Gel, hPL CM, and FBS control, respectively. Cells in hPL gel showed a 50% higher proliferation rate of viable cells compared to other culture media. AO/EtBr staining with FBS CM, hPL CM, and hPL gel revealed an increase in viable cells and a decrease in early apoptotic and necrotic cells in hPL Gel. In conclusion, the results of this study highlight the potential of hPL-based gels as superior matrices for multi-layered and enhanced proliferation of hUCT-MSCs.

Arginine methylation, catalyzed by protein arginine methyltransferases (PRMTs), is a regulatory key mechanism involved in various cellular processes such as gene expression, RNA processing, DNA damage repair. Increasing evidence highlights the crucial role of PRMTs in human diseases, including cancer, cardiovascular and metabolic diseases. Here, this review focuses on the latest findings regarding PRMTs in the central nervous system (CNS), emphasizing their regulatory roles in neural stem cells, neurons, and glial cells. Additionally, we examine the connection between PRMTs dysregulation and neurological diseases affecting the CNS, including brain tumors, neurodegenerative diseases, and neurodevelopmental disorders. Therefore, this review aims to deepen our understanding of PRMTs-mediated arginine methylation in CNS and open avenues for developing novel therapeutic strategies for neurological diseases.

This study was conducted to evaluate the impact of sodium hydrogen sulfide (NaHS) on the therapeutic efficacy of bone marrow mesenchymal stem cells (BM-MSCs) in the treatment of collagen-induced arthritis (CIA) rats. MSCs were isolated and cultured from rat bone marrow, and their characteristics were determined. The CIA model was induced in rats by intradermal injections of type II collagen on days 0 and 21. A variety of treatments were administered, including naproxen, BM-MSCs, BM-MSC-conditioned media, NaHS, BM-MSCs preconditioned with NaHS, and BM-MSCs preconditioned with NaHS-conditioned media. The infused BM-MSCs homed to the bone trabeculae and cartilage of the knee joint, leading to significant improvements in gait scores and a reduction in paw withdrawal frequency (PWF). Treatment with BM-MSCs and NaHS also significantly suppressed serum levels of CRP, RF, and 14-3-3η, while downregulating TNF-α gene expression and MMP-1 protein levels in the synovial membrane. Histopathological analysis confirmed these biochemical and molecular genetic findings. Notably, CIA rats treated with BM-MSCs preconditioned with NaHS showed the most significant improvements, with outcomes closely resembling those of healthy controls. This study concludes that NaHS enhances the therapeutic efficacy of BM-MSC therapy for rheumatoid arthritis (RA) by augmenting their anti-inflammatory, immunomodulatory, and regenerative properties.

Immune regulation is recognized as a cornerstone therapeutic strategy for the treatment of various autoimmune diseases. These disorders, driven by dysregulated immune responses, contribute significantly to morbidity and mortality. Although conventional immunosuppressive therapies provide symptomatic relief, their prolonged use is often associated with severe adverse effects, underscoring the need for safer and more effective treatment approaches. Extracellular vesicles (EVs), derived from immunoregulatory cells such as regulatory T cells, dendritic cells, mesenchymal stem cells, and neutrophils, have emerged as promising candidates for targeted immunomodulation. These nanoscale vesicles inherit the immunosuppressive properties of their parental cells, thereby facilitating immune homeostasis while mitigating the risks associated with other cell-based therapies. This review provides a comprehensive overview of recent advances in the application of immunoregulatory cell-derived EVs for autoimmune disease treatment, with a particular focus on their mechanisms of action within the immune microenvironment. Finally, we discuss the challenges and potential future directions in the development of EV-based therapies for autoimmune diseases.

Cryopreservation is an essential step for autologous hematopoietic stem cell (HSC) transplantation and umbilical cord blood units (CBUs), and for allogeneic peripheral blood stem cells (PBSCs) or bone marrow (BM) when immediate infusion is not possible. However, the cryoprotectant dimethyl sulfoxide (DMSO) used for HSC cryopreservation can be toxic to cells post-thaw and to patients during infusion. The Rubinstein solution is validated to wash HSCs, but the unavailability of Dextran40 in Italy prompted a search for alternatives. This report discusses the use of Gelofusine, a 4% modified gelatin solution, as a substitute for Dextran40-based solutions in washing cryopreserved stem cell products.

Nonsense-mediated RNA decay (NMD) is a highly conserved RNA turnover pathway that influences several biological processes. Specific features in messenger RNAs (mRNAs) have been found to trigger decay by NMD, leading to the assumption that NMD sensitivity is an intrinsic quality of a given transcript. Here, we provide evidence that, instead, an overriding factor dictating NMD sensitivity is the cell environment. Using several genome-wide techniques to detect NMD-target mRNAs, we find that hundreds of mRNAs are sensitized to NMD as human embryonic stem cells progress to form neural progenitor cells. Another class of mRNAs escape from NMD during this developmental progression. We show that the differential sensitivity to NMD extends to in vivo scenarios, and that the RNA-binding protein, HNRNPL, has a role in cell type-specific NMD. We also addressed another issue in the field-whether NMD factors are core or branch-specific in their action. Surprisingly, we found that UPF3B, an NMD factor critical for the nervous system, shares only 30% of NMD-target transcripts with the core NMD factor UPF2. Together, our findings have implications for how NMD is defined and measured, how NMD acts in different biological contexts, and how different NMD branches influence human diseases.

The nucleolus is a membrane-less subnuclear compartment known for its role in ribosome biogenesis. However, emerging evidence suggests that nucleolar function extends beyond ribosome production and is particularly important during mammalian development. Nucleoli are dynamically reprogrammed post-fertilisation: totipotent early mouse embryos display non-canonical, immature nucleolar precursor bodies, and their remodelling to mature nucleoli is essential for the totipotency-to-pluripotency transition. Mounting evidence also links nucleolar disruption to various pathologies, including embryonic lethality in mouse mutants for nucleolar factors, human developmental disorders and observations of nucleolar changes in disease states. As well as its role in ribogenesis, new findings point to the nucleolus as an essential regulator of genome organisation and heterochromatin formation. This Review summarises the varied roles of nucleoli in development, primarily in mammals, highlighting the importance of nucleolar chromatin for genome regulation, and introduces new techniques for exploring nucleolar function.

Chronic myelomonocytic leukemia (CMML) is a hematologic malignancy with a poor prognosis and limited targeted therapies. Lysine demethylase 6A (KDM6A), a H3K27 demethylase and key component of the COMPASS complex, is frequently mutated in hematologic malignancies, but its roles in embryonic hematopoiesis and tumor suppression in CMML remain unclear. Using zebrafish models with kdm6a mutants and integrative multi-omics analysis (ATAC-seq, RNA-seq, ChIP), we find that Kdm6a is a critical positive regulator of hematopoietic stem and progenitor cell (HSPC) emergence via Syk-related inflammatory signaling in a H3K27me3-dependent manner. We further find that Kdm6a haploinsufficiency in zebrafish leads to myeloid-biased hematopoiesis and a CMML-like disease, similar to CMML patients with reduced KDM6A expression. This KDM6A haploinsufficiency also significantly alters the chromatin landscape of genes associated with aging and cellular homeostasis in HSPCs. Mechanistically, KAM6A haploinsufficiency represses SOCS3 expression, thereby activating JAK/STAT3 signaling in HSPCs. Importantly, inhibitors targeting JAK or STAT3 phosphorylation alleviate myeloid expansion, providing a rationale for JAK/STAT pathway inhibition in CMML therapy. These findings enhance our understanding of CMML pathogenesis and propose new therapeutic avenues.

This study have previously reported that ZIF-based chiral nanomedicines achieve Parkinson's disease (PD) therapy through differential metabolism and relief of neuroinflammation. However, lack of overall chirality and anti-inflammatory capacity of nanomedicines limit the further effective solution to the nanobiological effects research in PD. Here, it dexterously loaded chiral gold nanoclusters (AuNCs) onto the inner and outer surfaces of ZIF to achieve the purpose of simultaneously improving the overall chirality and anti-inflammatory activity of the composite nanoparticles (NPs). There are significant differences in the composition of protein corona between different chiral NPs, which elucidates the mechanism of chiral-mediated discrepancies in metabolism and the blood-brain barrier (BBB) traversing. Multi-omics and biochemical techniques further reveal that chiral NPs interfere with the chemokine axis (CX3CL1/CX3CR1)-NF-κB-NLRP3 and PI3K-AKT signaling pathways, regulate communications between neurons, neural stem cells and microglia ("the three-body problem"), and induce anti-inflammatory efficacy of microglia mitochondrial energy metabolic reprogramming in PD. The research uncovers the biodistribution, metabolic variances, and therapeutic mechanism of chiral NPs, providing deep insights into the nanobiological effects of chiral anti-inflammatory nanomedicines in PD therapy for future clinical transformation.

This work presents a strategy for generating composite hydrogels bearing photoconductive conduits held by supramolecular interactions that are compatible with digital light processing (DLP) printing. Conductive polymers are typically processed with organic solvents as the film, yet if used as biomaterials, excitable cells often require matching with the mechanical and structural properties of their native, aqueous three-dimensional (3-D) microenvironment. Here, we utilize peptide-functionalized porphyrin units capable of self-assembling into photoconductive nanostructures with defined nanomorphologies under aqueous conditions. In addition to the DXXD peptide arms (X = V, F), the sequence variants studied here include a peptidic moiety bearing allyloxycarbonyl (alloc) groups that can serve as crosslinking sites of the acrylate-based monomers that ultimately form the base 3-D covalent network for the hydrogels. We investigate the impact of pre-templating polymeric gelators with supramolecular assemblies vs. printing a dispersed peptide-porphyrin in a polymer composite, specifically, the potential impact of the morphologies of the supramolecular additives or "dopants" on the resulting mechanical property, conductivity, and printability of the hydrogels, comprised of a hybrid between acrylated polymers and supramolecular peptide-porphyrin assemblies. Lastly, we demonstrate the role of photophysical properties that emerge from peptide-tuned porphyrin assemblies as a photoabsorber additive that influences the printing outcomes of the composite hydrogel. Overall, we present a covalent-supramolecular composite hydrogelator system where the self-assembled networks offer a pathway for energy transport and mechanical reinforcement/dissipation at the same time, leading to the formation of a hydrogel with optoelectronic, mechanical, and printable behavior that can be influenced by self-assembled dopants.

Loss of moisture is the primary cause of skin ageing and dysfunction. The skin's hydration largely depends on hyaluronan (HA) and its ability to retain water. Ultraviolet (UV) irradiation, which accounts for 80% of skin ageing (commonly referred to as photoaging), gradually disrupts the balance of HA metabolism, leading to a reduction in HA levels, dehydration, and, ultimately, the formation of wrinkles.