Neural organoids can exhibit variability in both tissue shape and tissue identity. Here, we present a pipeline for rapid, protocol-agnostic quality control screening of brain organoids based on their overall gross morphology. We describe a semi-automated image analysis of organoid size, shape, and texture from 2D bright-field imaging. We provide a reference dataset of brain organoids with complex morphology. We show how to integrate input and reference organoids and perform the unbiased sample selection by k-means clustering. For complete details on the use and execution of this protocol, please refer to Chiaradia et al.1.

Knee osteoarthritis (OA) causes significant chronic pain and disability. Current non-operative treatments are largely symptom-modifying. While intra-articular mesenchymal stem cell (MSC) therapies are promising, randomized controlled trials (RCTs) report inconsistent results due to heterogeneity in cell sources, preparations, and techniques.

To assess the effectiveness of cord blood nucleated cell extract (CBNCE) in rheumatoid arthritis (RA) and explore the mechanism preliminarily.

Encapsulation of human pluripotent stem cells (hPSC) has high relevance for biomedical applications ranging from tissue engineering to drug screening and cellular therapies. Microcapsules serve a multitude of purposes-from promoting hPSC organization into spheroids to enabling scalable differentiation in vitro and providing immunoisolation in vivo. While multiple encapsulation strategies have been reported, there is typically a tradeoff between the structural complexity of capsules and throughput of their generation. Our paper describes a novel 3D printed microfluidic encapsulation device for fabricating structurally complex microcapsules with a hydrogel shell and aqueous core at rates of up to 1825 Hz. The use of 3D printing allowed to dramatically decrease device footprint when compared to standard soft lithography-based microfluidic encapsulation devices. Leveraging decreased footprint and the ability to arrange fluidic networks in a 3D space, we fabricated a 10-nozzle microencapsulation device that generated microcapsules at a rate 10-times that of the single-nozzle device. The novel microfluidic device was used to encapsulate hPSCs, human embryonic stem cells (hESCs), and induced pluripotent stem cells (iPSCs). Hydrogel microcapsules with aqueous core promoted hPSCs aggregation into spheroids or embryoid bodies, which maintained high levels of pluripotency. Our technology enables safe and effective encapsulation of hPSCs on a scale (billions of cells) required to treat patients.

The proliferative capacity of adipose tissue stem cells (ATSCs) declines with aging due to the onset of senescence. This study aimed to determine whether fractionated electron irradiation can induce stable senescence in human ATSCs.

Immune reconstitution (IR), acute graft-versus-host disease (aGVHD), and cytomegalovirus (CMV) infection are all crucial factors influencing the prognosis following hematopoietic stem cell transplantation (HSCT).

Craniofacial bone reconstruction presents significant clinical challenges due to the region's complex anatomy and the need to restore both structural integrity and aesthetic function. This study aimed to enhance bone regeneration at graft sites through the development of an osteoconductive bioactive glass-polymer composite for patient-specific implant (PSI) applications. Composite constructs were fabricated via extrusion-based 3D printing using varying weight ratios of bioactive glass ceramic (BGS-7) and poly(ε-caprolactone) (PCL): 2:8, 4:6, 4.5:5.5, and 5:5. Printing parameters were optimized for each formulation to ensure consistent material flow and structural fidelity. Comprehensive characterization included morphological and elemental analysis via scanning electron microscopy/energy-dispersive X-ray spectroscopy, mechanical testing (compressive, flexural, and tensile), and biological evaluation and osteogenic differentiation of human placental stem cells. Higher BGS-7 content correlated with increased incorporation of phosphate, silicon, and calcium, contributing to enhanced mechanical properties and osteogenic potential. The constructs supported high cell viability, promoted cell adhesion and spreading, and induced osteogenic differentiation, as evidenced by calcium deposition and upregulation of key markers. The high-content BGS-7/PCL (5:5) formulation demonstrated optimal printability and bioactivity and was successfully used to fabricate anatomically accurate, human-scale structure. These findings highlight the potential of BGS-7/PCL composites as scalable, biocompatible, and osteoconductive platforms for craniofacial PSI applications.

Mutations in the presenilin (PS/PSEN) genes cause early-onset familial Alzheimer's disease (AD) by enhancing cerebral accumulation of amyloid-β (Aβ) peptides and microtubule-associated protein tau (MAPT). How PS mutations affect Aβ generation is well characterized, but the precise cellular mechanisms by which PS dysfunction drives neuronal tau pathology are not fully understood. Here, we investigated the mechanisms linking PS/γ-secretase-dependent tau pathology and autophagy/proteasome by employing pathological, imaging and molecular approaches in human brains, fibroblasts and induced pluripotent stem cells (iPSC)-derived neurons from PSEN1-linked familial AD carriers, and in a novel neuronal PS-deficient tauopathy transgenic mouse. We found enhanced levels and colocalization of pathological phosphorylated tau (pTau) and ubiquitin factor p62 in the hippocampus of dementia patients with familial AD-linked PSEN1 mutations, corticobasal degeneration and Pick's disease, suggesting disrupted proteasomal degradation in tauopathies. Human primary fibroblasts from PSEN1 G206D and/or L286P carriers showed elevated LC3-I and autolysosomes indicating autophagy flux alterations. Human iPSC-derived neurons harboring the familial-AD linked PSEN1 G206D mutation showed increased aggregated tau and reduced secreted tau, whereas pharmacological proteasome inhibition reduced significantly total and pTau (Ser396/404) while increasing its release. Consistently, proteasomal inhibition decreased intracellular tau and pTau and promoted tau release in human tau-expressing neurons through a mechanism that partially depends on PS. In the hippocampus of neuronal PS-deficient mice, Akt activation and GSK3β inhibition were associated with elevated levels of phosphorylated and aggregated tau and the ubiquitin-binding protein p62. In conclusion, PS function is required for autophagy/proteasome-mediated tau elimination in neurons, whereas that FAD-linked PSEN1 mutations cause progressive tau pathology by disrupting the proteasome and autophagy/lysosomal pathways.

Periodontitis can impair the osteogenic function of periodontal ligament stem cells (PDLSCs), thereby compromising their capacity for periodontal tissue regeneration. In this study, we explored the impact of a synthetic small molecule, DS96432529 (DS), on the osteogenic differentiation potential of PDLSCs and its underlying mechanism.

Mesenchymal stromal cells (MSCs) are widely used in regenerative medicine, but their clinical utility is limited by replicative senescence. Strategies that reverse aging while maintaining MSC identity are urgently needed.

Diabetic retinopathy (DR), the most prevalent ocular complication of diabetes, progresses from non-proliferative (NPDR) to sight-threatening proliferative (PDR) stages. Current interventions-including retinal photocoagulation, intravitreal anti-VEGF agents, and surgery-address advanced disease, often require repeated administration, and carry risks like retinal injury. Safer, more effective, and longer-lasting treatments are needed, especially for early-stage DR. Mesenchymal stem cells (MSCs) and their derivatives offer a promising alternative, with advantages including low immunogenicity, paracrine signaling, and the ability to mitigate inflammation and vascular permeability. However, challenges in delivery efficiency and targeting specificity remain. Hydrogel-based scaffold materials are increasingly important due to their superior biocompatibility and ability to overcome ocular barriers. Recent advances include novel injectable hydrogels that can be combined with drugs or stem cells, enabling targeted delivery to retinal layers, prolonging therapeutic retention, and significantly improving bioavailability for sustained treatment of DR.

Fatty acid (FA) oxidation plays an important role in T cell responses. However, whether DGAT1-mediated FA esterification to triacylglycerol also regulates T cell function remains unclear. Here we uncover a sexually dimorphic requirement for DGAT1 expression in CD8+ tumour-infiltrating lymphocyte function. In female mice, T cell-specific Dgat1 deficiency improves mitochondrial metabolic fitness and expands the pool of progenitor exhausted CD8+ T (Tex) cells to sustain antitumour responses. In male mice, however, Dgat1 deficiency leads to FA peroxidation, endoplasmic reticulum (ER) stress and CD8+ Tex cell death. We show that these effects are mediated by androgen receptor (AR) signalling. Deletion of Ar, overexpression of glutathione peroxidase 4, or inhibition of ER stress-induced cell death rescues Dgat1-deficient CD8+ T cell survival and promotes antitumour responses in male mice. Overall, this study suggests that DGAT1 detoxifies AR signalling in male mice to protect against ER stress-induced cell death and maintain T cell stemness, and uncovers sex-specific metabolic adaptations in the tumour microenvironment.

Sex chromosome loss (SCL) occasionally occurs after sex-mismatched allogeneic hematopoietic stem cell transplantation (allo-HSCT), but its clinical significance remains unclear. This retrospective study analyzed 78 patients who developed SCL after sex-mismatched HSCT, comprising 27 cases with initial-onset SCL (detected within first month post-transplant) and 51 cases with late-onset SCL (median onset time: 6.1 months; range: 2-73.1 months). Based on the results of chimerism analysis, initial-onset SCL was found to predominantly reflect the physiological aging hematopoietic cells from older donors, rather than relapse-related clonal abnormalities. In contrast, late-onset SCL was significantly associated with worse overall survival (OS) (HR 8.190,95% CI 2.842-23.600), inferior event-free survival (EFS) (HR 4.691,95% CI 1.839-11.966), and higher relapse risk (HR 6.751,95% CI 1.912-23.841). Among late-onset SCL patients, the optimal initial SCL clone size threshold for predicting relapse was 6% (sensitivity of 88.6%, specificity of 75.0%). The multivariate analysis confirmed ≥6% initial SCL clone size as an independent risk factor for relapse (HR 3.546, 95%CI 1.540-8.161), EFS(HR 3.418, 95% CI 1.572-7.433) and OS (HR 11.665, 95% CI 3.926-34.663). In conclusion, late-onset SCL may be associated with poor clinical outcomes, with the ≥6% clonal threshold serving as a critical marker for identifying high-risk patients prone to treatment failure.

Diabetic encephalopathy (DE) is a prevalent complication of diabetes which can lead to cognitive dysfunction, without effective therapy currently. In diabetic patients, a reduction in adult hippocampal neurogenesis (AHN) is a heightened risk of cognitive impairment, which may be associated with neuroinflammation caused by microglia. In this study, we established a DE mouse model and conducted in vitro cultures of microglial cells and neural stem cells. Our study demonstrated that the high-glucose associated with DE impairs AHN and induces microglial NOD-, LRR-, and pyrin domain-containing protein 3 (NLRP3) dependent pyroptosis. Further investigation showed that upregulation of microglial NLRP3 promotes the activation of Gasdermin D (GSDMD), the key pyroptosis effector, and the cleavage of pro-interleukin-1β (pro-IL-1β) by caspase-1, exacerbated pyroptosis and induced release of IL-1β, which might lead to impaired AHN and subsequent cognitive dysfunction. Conversely, downregulation of microglial NLRP3 inhibited caspase-1 activation and pyroptosis, reduced release of IL-1β, improved AHN, and rescued cognitive deficits in DE mouse model. Such findings suggest that targeting microglial NLRP3 inflammasome-mediated pyroptosis may be an important potential therapeutic target for treating DE.

The activation of the embryonic genome is a crucial step in development. In addition to thousands of genes, many transposable elements (TEs) are robustly transcribed during early mammalian development. However, their transcriptional regulators remain largely unexplored. Here, we set out to identify transcription factors regulating the expression of TEs from the LINE, SINE and ERVL families during mouse preimplantation development. In particular, the MaLR family are the most abundant ERVL in the mouse genome and are also the most abundant constituent of the transcriptome in early mouse embryos. We find that the general transcription factor TBP binds and activates MaLRs in mouse embryos. Loss-of-function of TBP leads to downregulation of MaLRs, specifically the ORR1A family, which is the youngest ORR subclass and contributes a significant portion of major zygotic genome activation transcripts. Our work identifies regulators of TE expression in vivo and highlights a previously unrecognised role for the general transcription factor TBP in regulating a highly specific TE transcriptional programme.

Microfragmented adipose tissue (MFAT) has emerged as a minimally manipulated, autologous orthobiologic for musculoskeletal disorders, with the Lipogems system representing one of the most widely studied platforms. By preserving the stromal vascular niche and pericyte-rich microenvironment, MFAT provides a ready-to-use biologic without enzymatic digestion or ex vivo expansion. This narrative review synthesizes the biological rationale, clinical applications, and comparative effectiveness of MFAT across major orthopedic indications. We review action mechanisms, patient selection factors, procedural considerations, and regulatory context, with emphasis on outcomes, durability, and imaging correlations. Across joints, MFAT has demonstrated consistent improvements in pain and function, particularly in early to moderate osteoarthritis. However, radiographic findings are heterogeneous and often discordant with clinical outcomes. Studies suggest MFAT offers outcomes comparable to platelet-rich plasma and bone marrow aspirate concentrate rather than clear superiority. Overall, the evidence base is dominated by low- to moderate-level studies with small sample sizes, protocol variability, and limited follow-up. This review provides a cross-joint clinical synthesis to contextualize MFAT's current role in orthopedic practice, highlighting its procedural simplicity and favorable safety profile. Despite encouraging clinical signals, we strongly underscore the need for future high-quality trials, standardized methodologies, and cost-effectiveness analyses to define its long-term clinical utility.

The Fat gene family, also known as atypical cadherins, contributes to the formation of planar cell polarity, which determines organ shape, size, and polarity. However, their roles in tooth morphogenesis remain unclear. In this study, the aim was to investigate the spatiotemporal expression patterns of Fat1-Fat4 during murine tooth development.

Embryonic regulators are often re-expressed in cancers; however, the functional and molecular significance of this is not always understood. The epigenetic priming factors developmental pluripotency-associated 2 and 4 (DPPA2/4) have crucial roles in early development and are implicated in cancer pathogenesis. We reveal that in non-small cell lung cancer (NSCLC), DPPA2/4 coexpression is associated with poorly differentiated tumors and impaired patient outcomes. Biochemically, human DPPA2/4 multimerize for their protein stability and enhanced nucleosome binding activity. In NSCLC cells, DPPA2/4 bind CG-rich sequences including promoters of developmental genes, Wnt signaling, and catabolic genes. Chromatin state modeling revealed that DPPA2/4 preferentially bind active H3K4me3 and H3K27ac domains that were intriguingly also enriched for PRC1 and its product, H2AK119ub, which was validated by H3K4me3-H2AK119ub sequential ChIP. Knockdown experiments revealed that DPPA2/4 were required to maintain RING1B and H2AK119ub at these domains. Surprisingly, despite the presence of PRC2.1, these regions lacked any detectable H3K27me3, suggesting an uncoupling between the recruitment of PRC2 to chromatin and its catalytic product. When exogenously overexpressed in NSCLC cells where they are not normally present, DPPA2/4 bind to and promote active chromatin states, resulting in an increase in in vivo xenograft tumor growth. Our results demonstrate how, in NSCLC cells, DPPA2/4 act as molecular amplifiers of active and poised chromatin. Together, this highlights how aberrant reactivation of embryonic factors in cancers may take on new functions, promoting tumorigenesis.