Chronic inflammation is increasingly recognized as a key factor in the progression of diabetic kidney disease (DKD). By discovering that the regulation of gut microbiota plays an important role in diabetic kidney disease, human umbilical cord mesenchymal stem cells (HU-MSCs) explore the mechanism of fibrosis in diabetic kidney disease through the regulation of chronic inflammation, providing new clinical insights for the prediction, diagnosis, and treatment of diabetic kidney disease.

Montrichardia linifera (Arruda) Schott (M. linifera) is commonly used by Amazonian riverine communities for the treatment of skin ulcers, although its effects as a wound healer have never been evaluated until now. Therefore, the in vitro wound-healing activity of the extracts from the stem and petiole of M. linifera was investigated for the first time.

Cardiac organoid is a miniature and simplified three-dimensional (3D) cellular model system grown from progenitor cells or stem cells that more accurately mimic the significant biological characteristics and functions of the normal cardiac system than conventional two-dimensional (2D) models. With continued advances in 3D culture approaches, the cardiac organoid models produced through self-organization strategy following developmental induction conditions exhibit higher metabolic similarities and physiological relevance. Increasing evidence demonstrates that cardiac organoids based on the in vitro model system are useful platforms for studying human cardiac biology and pathophysiology. Despite significant advancements, the development of cardiac organoids has not progressed as far as other types of organoids due to the intricate cellular structure and microenvironment of the heart. In this review, we highlight the current classification and bioengineering strategies for establishing cardiac organoids using Matrigel and decellularized extracellular matrix derived culture platforms followed by a review of contemporary reports of their use in development biology, disease modeling, drug testing and efficacy evaluation. We also shed the light in the current limitations and future perspective of the cardiac organoid to motivate future research and accelerate the widespread adoption of organoids platforms.

Adipose-derived stem cells (ADSCs) have shown great potential in cartilage tissue engineering (TE) due to their accessibility, high proliferation rate, and chondrogenic differentiation capacity. This study aims to systematically evaluate research trends, collaboration patterns, and emerging themes in ADSC-based cartilage TE through a bibliometric analysis, providing actionable insights to address knowledge gaps and advance the field.

Chimeric antigen receptor (CAR) T cells are a leading immunotherapy for refractory B-cell malignancies, but their impact is constrained by toxicity and incomplete long-term efficacy. Daily (circadian) rhythms in immune function may offer a lever to boost therapeutic success. Studies suggest that time of day influences immune-based therapies, including vaccines, hematopoietic stem cell transplantation, and checkpoint inhibitors for cancer. However, the clinical relevance of biological rhythms to CAR-T cell therapy remains unknown.

Fertility preservation (FP) includes all the methods to preserve germ cells, reproductive tissues, or embryos for the future reproduction of patients at risk of infertility. Cryopreservation is an essential step of FP, storing the specimens in subzero temperatures to suppress cellular metabolism and restore cryopreserved specimens for future use. Although oocyte cryopreservation (OC) and embryo cryopreservation (EC) are two accepted methods of FP in women, ovarian tissue cryopreservation (OTC) is a novel method that is favorable in patients who are not appropriate candidates for OC and EC, and those who suffer from irritating menopause symptoms caused by estradiol deficiency. OTC has shown promising results in restoring fertility and the endocrine function of ovaries. Slow freezing and vitrification are two well-established methods for cryopreservation of biological specimens. Despite recent developments in the vitrification of ovarian tissue and comparable results to slow freezing, we lack a standard protocol for ovarian tissue vitrification, and slow freezing is still the preferred method in most centers. Under an acceptable medical condition and desirability, transplantation of cryopreserved tissue is performed either in orthotopic sites (orthotopic transplantation, OT) such as the original site of the ovaries and uterus, or heterotopic sites (heterotopic transplantation, HT) like abdominal wall, forearm, and peritoneal lining. Although both sites of transplantation are associated with endocrine function recovery, OT better restores fertility. This review will focus on OTC and its types, ovarian tissue transplantation, and efficacy in clinical practice.

Aging is associated with a decline in the function of hematopoietic stem cells (HSCs). This decline in HSC function results in reduced hematologic regenerative capacity and an increased incidence of hematologic disorders. In general, aged HSCs show on average an increase in cell size and a lower frequency of cells polar for protein polarity markers. The size of an HSCs has been proposed to be tightly linked to the potential of the HSCs, with small HSCs showing a higher potential compared to large HSCs. The increase in size of HSCs upon aging may be associated with the reduced potential of aged HSCs. HSCs are located within the bone marrow (BM) in distinct microenvironments called niches. These niches provide critical physical and molecular signals that are essential for HSC self-renewal, proliferation, migration and differentiation. There are multiple types of functional niches, and HSCs within these distinct types of niches show a distinct type of potential. Furthermore, the distribution of HSCs relative to niches changes upon aging. It is not known whether there is a correlation of HSCs size, HSCs polarity and the location of HSCs in distinct types of niches, as might be expected, as all three (size, polarity and position) have been linked to HSC potential. Here we show that in young mice smaller HSCs, which are more myeloid-biased, are preferentially located at central BM niches, including sinusoids and megakaryocytes. In contrast, larger HSCs, which show a bias toward B-lymphoid differentiation, are preferentially located in endosteal BM niches close to arterioles. However, in aged mice, which also contain HSCs of different sizes, there was no correlation between HSC size and localization and potential. Furthermore, within the hematopoietic stem and progenitor cell (HSPC) population, cell size increases as the cells become more limited in their capacity. Notably, we further report that changes in the level of polarity correlate with HSC potential even in aged mice.

Percutaneous coronary intervention (PCI) remains the primary treatment for coronary artery disease (CAD), yet post-procedural arterial injury triggers cellular change and pathological inflammation, leading to thrombosis and restenosis. Recent studies have highlighted the chemokine CXCL2 play an important role in the immune response to tissue repair. However, the cellular mechanisms and the role of chemokine CXCL2 underlying arterial repair after PCI remain poorly understood.

Follicle Stem Cells (FSCs) in the germarium of a Drosophila melanogaster ovary are maintained through independent regulation of division and differentiation. Adult FSCs can become proliferative Follicle Cells (FCs) to the posterior, or quiescent Escort Cells (ECs) to the anterior. Graded extracellular Hedgehog (Hh) and Wnt signals emanate from cells anterior to FSCs (Cap Cells and ECs) to guide these behaviors together with an inverse JAK-STAT pathway stimulated by ligand from a more posterior source (polar FCs). Here we used lineage analyses to investigate the role of those signals in the development of ECs, FSCs and FCs from a common set of precursors during pupation. Previous studies found that the most anterior precursors divide slowest, with quiescence spreading to all future ECs from the anterior, FSCs are specified simply by their location at eclosion, and the first FCs derive from a group of cells that accumulates posterior to the developing germline over the first 48h of pupation. We now show that the latter cells derive from migration of precursors out of the developing germarium. We also found that Wnt pathway activity favored conversion of precursors to more anterior adult derivatives, while JAK-STAT pathway activity favored posterior outcomes. Wnt pathway activity increased over the first 48h and maintained a graded pattern throughout pupation, terminating at the anterior extent of FC specification, commensurate with Wnt pathway activity continuously opposing FC formation. JAK-STAT pathway activity was consistently lowest in anterior cells, indicating a posterior source early in pupation prior to formation of the first polar FCs. Precursor division was promoted by JAK-STAT signaling and also by Hh signaling, acting through transcriptional induction of yorkie . Faster division favored a precursor becoming an FSC, as seen for FSC lineage maintenance in adults.

Subcellular proteomics holds the potential to reveal the molecular architecture of cellular processes with unprecedented spatial resolution. Performing these analyses deeply, at the level of hundreds to thousands of proteins in subcellular resolution, is a high and still unmet technical need. Here, we advance microprobe capillary electrophoresis-mass spectrometry (CE-MS) to achieve deep proteomic coverage-quantifying over 1,000 proteins within opposing poles of an asymmetrically dividing embryonic stem cell (blastomere). We integrated CE-electrospray ionization (CE-ESI) with trapped ion mobility spectrometry time-of-flight (timsTOF) MS, implementing data-independent acquisition (DIA) via parallel accumulation-serial fragmentation (diaPASEF). This CE-diaPASEF workflow identified 1,035 proteins from ∼200 pg of proteome digest, equivalent to ∼80% of the HeLa cell's content, with high reproducibility (coefficient of variation <15% across technical triplicates). With microprobe sampling, this technology quantified 808 to 1,022 proteins in opposing poles of a dorsal-animal (D1) blastomere in the 8-cell Xenopus laevis embryo. Comparative proteomic analysis of the D1 blastomere and its descendants-the dorsal-animal-midline (D11) and dorsal-animal-lateral (D12) cells-revealed diverse molecular outcomes of asymmetric division: some protein profiles remained conserved, while others underwent significant or even reversed changes as these lineages descended into neural tissue and epidermal trajectories. Ultraviolet light-induced ventralization was performed to help disentangle subcellular gradients from dorsal-ventral patterning. Collectively, this work establishes microprobe CE-diaPASEF as a powerful platform for deep subcellular proteomics, enabling new insights into spatial proteome organization during key developmental processes.

Comparative anatomical studies of primates and extinct hominins, including Neanderthals, show that the modern human brain is characterised by a disproportionately enlarged neocortex relative to the striatum. To explore the molecular basis of this difference, we screened for missense mutations that are unique to modern humans and occur at high frequency and that alter post-translational sites. One such mutation was identified in DCHS1 , a protocadherin family gene, and it was found to disrupt an N-glycosylation site in modern humans. Using CRISPR/Cas9-editing we introduced into human-induced pluripotent stem cells (hiPSCs) this ancestral DCHS1 variant present in Neanderthals and other primates, representing the ancestral state before the modern human-specific substitution. Leveraging hiPSCs-derived neural organoids, we observed an expansion of striatal progenitors at the expense of the neocortex, mirroring the anatomical distribution seen in non-human primates. We further identify the ephrin receptor EPHA4 as a binding partner of DCHS1 and show that modern human-specific alterations in DCHS1 modulate EPHA4-ephrin signalling, contributing to a gradual shift in the neocortex-to-striatum ratio - a hallmark of brain organisation in our species.

Genome maintenance is of the utmost importance in stem cells, as mutations can be propagated and cause defects in derivative tissues. Many stem cell types display low mutation rates, with embryonic stem cells (ESCs) being a notable example. The bases for this property are unclear but may be achieved by optimization of various processes including high-fidelity DNA repair, cell cycle checkpoint controls, and hypersensitivity to genotoxic insults that trigger cell death. Here, we investigate the mechanisms underlying the unique responses of mouse ESCs (mESCs) to replication stress (RS) using an array of small molecule inhibitors and genotoxins. We find that whereas mESCs survive under acute RS in an ATR- and CHK1-dependent manner similar to somatic cells, they lack a strong G2/M checkpoint and fail to repair DNA crosslinks in the absence of ATR signaling. Despite the lack of a strong G2/M checkpoint, mESCs maintain a spindle assembly checkpoint (SAC). We posit that mESCs preferentially repair DNA crosslinks in S phase via homology-directed mechanisms, and cells that fail to complete repair before mitosis undergo mitotic catastrophe and cell death. These findings shed light on mutation avoidance mechanisms in ESCs that may extend to other stem cell types.

Tissue-resident macrophages (TRMs) are innate immune cells that participate in tissue development, homeostasis, and immune surveillance. Extensive efforts have been made to recapitulate TRM development from pluripotent stem cells (PSCs) in vitro to study molecular and cellular mechanisms of TRM development and to create cellular models of disease. However, available PSC models of mouse TRM development exhibit low overall efficiencies of TRM generation, produce heterogeneous off-target populations, and rely upon undefined media components, thus limiting their reproducibility, scalability, and widespread application as an experimental platform for TRM biology. To address these important limitations, we developed an efficient and reproducible protocol to faithfully recapitulate the stepwise differentiation of mouse PSCs (epiblast stem cells) into unspecialized, proliferative TRMs through the pro-definitive hematopoietic program under defined conditions. These immature TRMs can stably integrate into developing mouse neural organoids in vitro and acquire features of microglia. In addition, PSC-derived immature TRMs can stably engraft into the lung niche in vivo and adopt alveolar macrophage characteristics. This new platform for modeling mouse TRM development represents a powerful experimental model system for studying TRM function and dysfunction in development and disease.

Elements such as iron, copper and zinc play essential roles in the mammalian oocyte, egg, and embryo, however among these metals, zinc plays unique regulatory roles. Temporal fluctuations in zinc concentrations drive reproductive milestones such as meiotic resumption, egg activation, and initiation of the mitotic cell cycle. Roles for zinc in late preimplantation embryo development, have not been well characterized. Using a quantitative element approach we report the inorganic profiles of mouse embryos progressing through the late blastocyst stage. We find that blastocysts, like oocytes and eggs, and distinct to somatic cells, maintain higher levels of zinc than copper and iron. All three of these essential metals are more abundant in the inner cell mass, which contains the population of pluripotent stem cells that give rise to the fetus, relative to the trophectoderm which gives rise to the placenta and extraembryonic tissues. To test whether zinc abundance was associated with mitotic progress and cell fate lineage, we perturbed zinc homeostasis during blastocyst formation by artificially raising intracellular zinc concentrations with zinc pyrithione. This treatment during the morula-to-blastocyst transition when cell fate lineages emerge resulted in an elevation of zinc in the ICM. This treatment did not impact cell number, but did increase expression of the pluripotency and epiblast marker, Nanog , and decreased expression of the primitive endoderm marker, Gata4 . These results demonstrate that the inorganic profiles of the late preimplantation embryo retain elemental hallmarks of earlier developmental stages and perturbation of zinc levels alters pluripotency gene expression in the blastocyst.

The presence of histone modifications associated with both transcriptional repression (H3K27me3) and activation (H3K4me3) on key developmental promoters in embryonic stem cells results from the co-localization of repressive Polycomb group (PcG) and activating Trithorax group (TrxG) protein complexes. Functional interactions between PcG and TrxG on these promoters are not fully understood. Here we focus on the relationships between Fbxl10/Kdm2b, a component of a PcG complex PRC1, and Kmt2b/Mll2, an essential component of TrxG at bivalent promoters. Computational analysis of previously published data revealed genome-wide correlation between chromatin occupancies of these two proteins, suggesting potential crosstalk between Kdm2b and Mll2 at both active and repressed promoters. We tested this hypothesis experimentally and found that loss of Kdm2b resulted in depletion of Mll2 at promoters genome-wide, suggesting that Kdm2b is required for Mll2 occupancy at both bivalent and active promoters. Loss of Kdm2b or the core PRC1 component Ring1b also resulted in the reduction of H3K4me3 specifically at bivalent promoters. These findings provide a direct pathway for cooperation between PcG and TrxG at bivalent promoters, suggesting an unexpected modification to the current model of bivalency. In addition, these findings reveal genome-wide role of Kdm2b independent of the full PRC1 complex.

Stem cells require signals from a cellular microenvironment known as the niche that regulates identity, location, and division of stem cells. Niche cell identity must be properly specified during development to form a tissue capable of functioning in the adult. Here, we show that the Tbx1 ortholog org1 is expressed in Drosophila testis niche cells in response to Slit and FGF signals. org1 is expressed during niche development and is required to specify niche cell identity. org1 mutants specified fewer niche cells, and those cells showed disruption of niche-specific markers, including loss of the niche adhesion protein Fas3 and reduced hedgehog expression. We found that org1 expression in somatic gonadal precursors is capable of inducing formation of additional niche cells. Disrupted niche identity in org1 mutants resulted in niche assembly and functionality defects. We find the conserved transcription factor islet is expressed in response to org1 and show that islet functions downstream to mediate niche identity and assembly. This work identifies a novel role for org1 in niche establishment.

In metazoans, nucleosomes harboring H3K79 methylation (H3K79me) deposited by the histone methyltransferase DOT1L decorate actively transcribed genes. Although DOT1L is implicated in transcription regulation and pathogenesis of human diseases such as leukemia and neurological disorders, the role of H3K79me in these biological processes remains elusive. Here, we reveal a novel functional synergism between H3K79me and H3K36 tri-methylation (H3K36me3), another histone modification enriched at active genes, in regulating gene expression and neural cell fate transition. Simultaneous catalytic inactivation of DOT1L and the H3K36 methyltransferase SETD2 via gene editing leads to the global loss of H3K79me and H3K36me3, hyperactive transcription, and failures in neural differentiation. Interestingly, the loss of H3K79me and H3K36me3 causes increased transcription elongation, gained chromatin accessibility at a group of enhancers, and increased binding of TEAD4 transcription factor and its co-activator YAP1 at these enhancers. Furthermore, YAP-TEAD inhibition partially restores the expression levels of hyperactivated genes upon H3K79me/H3K36me3 loss. Taken together, our study demonstrates a synergistic role of H3K79me and H3K36me3 in regulating transcription and cell fate transition, unveils novel mechanisms underlying such synergism, and provides insight into designing therapies that target diseases driven by misregulation or mutations of DOT1L and/or SETD2.

The dynamic regulation of epigenetic states relies on complex macromolecular interactions. PRC2, the methyltransferase complex responsible for depositing H3K27me3, interacts with distinct accessory proteins to form the mutually exclusive subcomplexes PHF1-PRC2.1, MTF2-PRC2.1, PHF19-PRC2.1, and PRC2.2. The functions of these subcomplexes are unclear and thought to be highly redundant. Here we show that PRC2 subcomplexes have distinct roles in epigenetic repression of lineage-specific genes and stem cell differentiation. Using a human pluripotent stem cell model, we engineered a comprehensive set of separation-of-function mutants to dissect the roles of individual protein-protein and DNA-protein interactions. Our results show that PRC2.1 and PRC2.2 deposit H3K27me3 locus-specifically, resulting in opposing outcomes in cardiomyocyte differentiation. We find that MTF2 stimulates PRC2.1-mediated repression in stem cells and cardiac differentiation through its interaction with DNA and H3K36me3, while PHF19 antagonizes it. Furthermore, MTF2-PRC2.1 maintains normal cardiomyocyte function. Together, these results reveal the importance and specificity of individual macromolecular interactions in Polycomb-mediated epigenetic repression in human stem cells and differentiation.

During embryogenesis, hematopoietic stem cells (HSC) and HSC-independent progenitor cells form concurrently and produce overlapping and distinct immune cell types. The full repertoire of immune cells generated during development and their origin across vertebrates is incompletely elucidated. Here, we performed temporal lineage tracing of the emerging hematopoietic system during zebrafish larval development followed by single cell RNA sequencing and identified a greater diversity of lymphoid cells than previously recognized. The cells are comprised of T-lymphocytes and Innate Lymphoid-like Cells (ILCs) that are Runx1-dependent. Both T-lymphocytes and ILC-like cells depended on Il2rγ, while only T-lymphocytes depended on Rag1. The larval ILC-like cells were detected in lymphoid and mucosal organs and were responsive to viral mimicry-induced stimulation, indicating their functionality in early vertebrate life. The work provides new fundamental knowledge on the early establishment of immune hierarchies and opens the zebrafish model to broader exploration of lymphoid immunity origination and function.

Pan-GSK3α/β inhibition promotes stem cell self-renewal through activation of WNT/β-catenin signaling, but its broad effects complicate the precise control of stem cell states. Here, we show that selective inhibition of GSK3α with BRD0705 supports the long-term self-renewal of mouse embryonic stem cells (ESCs), epiblast stem cells (EpiSCs), and neural stem cells (NSCs), independent of β-catenin signaling. When combined with the tankyrase inhibitor IWR1, BRD0705 broadly supports the maintenance of diverse pluripotent stem cell states, including ESCs, EpiSCs, and formative pluripotent stem cells. This BRD0705/IWR1 cocktail enables stable co-culture of naive ESCs and primed EpiSCs while preserving their distinct molecular and functional identities. Single-cell transcriptomics, epigenomic profiling, and functional assays confirm sustained lineage-specific features across stem cell types. These findings demonstrate that selective GSK3α inhibition enhances stemness by buffering against differentiation cues and promoting intrinsic self-renewal capacity. This work identifies GSK3α as a key regulator of self-renewal across distinct stem cell states and establishes a versatile culture system with broad applications.