The functional versatility of adipose tissue is mediated by various cell types and their capacity to transition between distinct functional states upon pathophysiological stimuli. Recent advances in single-cell transcriptomics have enabled the identification of novel and numerous cell types and states of adipocytes, adipose stem and progenitor cells, immune cells, vascular cells, and mesothelial cells. Significant efforts have been made to understand the key features and underlying processes of the cell subpopulations and to elucidate the cellular mechanisms of adipose tissue plasticity. In this review, we describe current knowledge of heterogeneous cell types and states in adipose tissue and emerging techniques to characterize them, offering fundamental knowledge for studying adipose tissue in the single-cell era.

Cultured meat has emerged as a promising approach to enhancing global food sustainability. Current production methods, however, rely mainly on single-cell-type culture, which fails to replicate the complex composition of conventional meat. In this study, a scaffold-free co-culture system was established to generate structured microtissue comprising highly differentiated muscle and fat cells. Co-cultured spheroids with the ratios of piscine adipose-derived stem cells (PADSCs) to piscine satellite cells (PSCs) being 5:5, 7:3 and 9:1 were investigated, and the result showed that all ratios remained robust and highly viable throughout the differentiation process and successfully induced the formation of bionic tissue containing both myotubes and adipocytes. Based on transcriptome data, the cluster analysis indicated that co-cultured spheroids (the ratio of PADSCs to PSCs was 7:3) were similar to the PADSC spheroids, and the expression levels of cell cycle-related and extracellular matrix-related genes were biased towards those of PADSC spheroids. These results indicated that these co-cultured spheroids held promise as building blocks for further tissue assembly and offered a promising approach for scaling up the production of structured cultured fish meat.

Neurogenesis, the creation of new neurons from neural stem cells (NSCs) in the brain, plays a crucial role in neurological diseases when disrupted. Herbal medicine components, especially those with dual applications in disease treatment and food, like those from Gastrodia elata Blume, have gained attention for their ability to influence neurogenesis. Notably, gastrodin and gastrodigenin from this herb influence neurogenesis and affect conditions like Alzheimer's, depression, stroke, and amnesia. Understanding these processes and mechanisms is essential for addressing neurological disorders. We also discuss gastrodin's potential in aiding peripheral nerve regeneration and its therapeutic effects on neurological diseases through neurogenesis regulation. This review offers insights into gastrodin's therapeutic potential, encouraging further research to boost its efficacy in neurological diseases.

Gastric cancer (GC) remains a major public health concern both in Taiwan and worldwide. While advances in public health have reduced its incidence rate, clinical outcomes of advanced GC remain suboptimal with current standard therapy. The Wnt/β-catenin signaling pathway is frequently up-regulated in GC, promoting tumor progression. This study investigated the anti-tumor effects of PKF118-310, a small molecule inhibitor of the β-catenin-TCF/LEF interaction, in GC cell lines and patient-derived models.

Cervical cancer mortality has since long been associated with HPV positivity. However, emerging evidence points to the significant role of cancer stem cells (CSCs) in tumorigenesis, metastasis, relapse, and chemoresistance. This necessitates targeting strategies not only against HPV+ cervical cancer cells but also against CSCs. Selective phosphodiesterase type 5 inhibitors are well-known as examples of successful drug repurposing. In a small screen, previous research identified cyclic guanosine monophosphate-phosphodiesterase (cGMP-PDE) inhibition as a potential strategy for targeting cancer stem cell maintenance and survival in various cell lines. Hence, herein we studied the status of cGMP-PDE and its modulation in cervical cancer.

Pancreatic ductal adenocarcinoma (PDAC) remains highly lethal due to gemcitabine (GEM) resistance. This study aimed to establish a clinically relevant immunocompetent model to identify novel mediators of acquired GEM resistance.

In our daily life, bone defects with sensory losing caused by limb trauma, degenerative diseases, or tumor resection are very common. When the bone defect size exceeds the critical size, it cannot spontaneously heal. Therefore, clinical intervention and artificial bone materials are needed to repair damaged bone tissue and restore sensation. Here, we reported that Collagen (Col) and Chondroitin sulfate (CS) adsorbed into hydroxyapatite (HA) for long-time sustained release was used to prepare nanofibrous scaffolds (Col+CS)@HA through electrospining, fibroblast growth factor (FGF) and CS + Col as two layers were used to prepare (Col+CS)@HA-(FGF/CS + Col)5 nanofiber scaffold through Layer-by-Layer (LBL) self-assembly technology. Interestingly, we found that neurogenesis could accelerate osteogenesis from RNA sequencing result. To confirm this result, we found that (Col+CS)@HA-(FGF/CS + Col)5 scaffold could promote osteogenesis process in bone mesenchymal stem cells (BMSCs) regulated through extracellular regulated protein kinases 1/2 (Erk1/2) activated runt-related transcription factor 2 (Runx2) pathway. Furthermore, the transcriptional levels of downstream genes associated with osteogenesis were significantly elevated. Intriguingly, we also found that (Col+CS)@HA-(FGF/CS + Col)5 scaffold could boost the differentiation of BMSCs into neurons and promote the transcriptional levels of neuron specific genes. As a result, we observed that new neuron was formed in Haversian canal in mice cranial defects area, which means (Col+CS)@HA-(FGF/CS + Col)5 scaffold could not only promote osteogenesis, but also could enhance neurogenesis. Therefore, we believe this study is promising, as it provides new insights towards the process of bone defect repairment along with sensation restorement.

T cell development in the thymus proceeds through coordinated transcriptional and spatial transitions. While key stages have been described, the precise timing and regulatory logic of lineage bifurcation-such as the separation between αβ and γδ T cells, and the commitment to CD4 or CD8 fate remains incompletely understood. A high-resolution, integrated view is needed to clarify how distinct T cell identities are established.

The regular emergence of influenza strains with pandemic potential necessitates vaccines that elicit protective immune responses across genetically diverse human populations. A critical but understudied factor is how germline-encoded variation in immunoglobulin genes shapes the development of neutralizing antibodies. Here, by combining personalized immunoglobulin genotyping with high-throughput paired-chain antibody sequencing from influenza A hemagglutinin (HA)-binding B cells across four donors, using a technique we developed called individualized single-cell analysis of paired expressed antigen receptors (ISCAPE), we demonstrate that B cell responses to HA are highly individual. We identified a common IGHV2-70 polymorphism that impaired the function of a class of neutralizing HA head-directed antibodies. Furthermore, we described HA central stem-targeting broadly neutralizing antibodies that utilize IGHD3-3 recombined with diverse IGHV genes, expanding the known repertoire of stem antibodies and highlighting antibody gene usage population restrictions. We suggest that multi-donor repertoire studies, coupled with personalized immunoglobulin genotyping, can uncover germline-encoded functional variations and help mitigate population vulnerabilities in vaccine design.

Autophagy is an essential developmental and homeostatic process, defined by the endolysosomal degradation of intracellular components and pathogens. Dysfunctional autophagy is implicated in complex human disease, yet reports of congenital autophagy disorders were considered exceedingly rare until the recent report of several unrelated families with bi-allelic variants in the core autophagy effector ATG7, complementing the report of two individuals harboring ATG5 variants. We now report six affected individuals from five families with bi-allelic ATG12 variants with complex neurological phenotypes overlapping those seen in individuals with pathogenic variants in ATG5 and ATG7: developmental delay, intellectual disability, congenital ataxia, hypotonia, and seizures with cerebellar vermis hypoplasia evident on neuroradiological imaging. Structural modeling implicated a potential disruption of the ATG12-ATG5-ATG16N-ATG3 complex. Biochemical analyses of primary fibroblasts confirmed the loss of stable ATG12-ATG5 conjugate in one family and altered autophagic flux in one unrelated family. The HaloTag processing assay in HeLa cells demonstrated a decrease in ATG12-ATG5 conjugate and reduced autophagic flux in response to starvation. Complementation studies demonstrated that equivalent missense atg12 variants were unable to fully recover the biochemical defect in atg12-null yeast, with microscopy analysis demonstrating a reduced delivery of autophagy substrates to the yeast's degradative compartment. Zebrafish studies confirmed that Atg12 is required for normal growth, brain development, and neural function. Collectively, our findings underscore the pivotal role of autophagy in maintaining human neural integrity, emphasize an emerging group of congenital autophagy disorders, and expand our understanding of adaptive homeostasis in human health and disease.

Dr. Shinya Yamanaka is recognized for the generation of induced pluripotent stem cells (iPSCs) from fibroblasts by a combination of multiple transcription factors, and he won the Nobel Prize in Physiology or Medicine in 2012 jointly with Sir John B. Gurdon for this discovery. Twenty years after the discovery, the Cell Reports Medicine editorial team discusses with Dr. Yamanaka the scientific, technical, and translational milestones that have shaped the field of regenerative medicine. We also discuss the role of iPSCs in disease modeling and drug discovery, the interplay with genome editing, and ongoing issues that still prevent the widespread clinical application of iPSC-derived therapies. Finally, Dr. Yamanaka reflects on promising, yet underexplored, applications of iPSCs.

Understanding biological processes requires spatiotemporal mapping of proliferative and transcriptional dynamics. Current spatial transcriptomics methods capture only protein-coding transcripts and static snapshots, obscuring non-coding RNAs (ncRNAs) and dynamic events. We developed SPTEdU-seq, integrating spatial total transcriptomics with 5-ethynyl-2'-deoxyuridine tracking to co-profile gene expression and proliferation dynamics. SPTEdU-seq demonstrates ultrahigh sensitivity for coding and non-coding transcripts and for splicing isoforms, with single-molecule probe design eliminating optical imaging. Applied to developing and adult mouse brains, it revealed spatial lncRNA patterns, reconstructed developmental trajectories, and enabled spatiotemporal lineage tracing. In murine ischemic stroke, it mapped regeneration dynamics and identified an Igfbp5+ astrocyte subtype within a pro-repair niche. In mouse and human renal tumors, it uncovered tumor-associated splicing and detected diagnostic 3p loss. By profiling newborn and resident cells in intact microenvironments, it unveiled previously inaccessible interaction networks. SPTEdU-seq thus establishes a powerful framework for investigating cell fate dynamics in regeneration, development, and cancer.

Tumors are increasingly recognized as a consequence of systemic immune dysregulation, while current therapies merely focus on direct tumor killing or local immune activation, overlooking the systemic immune landscape that enables tumorigenesis and metastasis. Targeting distal immune organs, such as the bone marrow (BM), without perturbing tumors remains challenging. Here, we develop a BM-targeted and tumor-evasive cell vector that restricts immunomodulation to the BM niche, enabling systemic immune reprogramming through niche-derived signaling. This mesenchymal stem cell (MSC)-based vector overexpresses Golgi apparatus protein 1 (MSCGlg1) to mimic BM affinity signals. In a myelosuppression model, MSCGlg1 delivers CDK4/6 inhibitors (CDK4/6i) to protect hematopoietic stem and progenitor cells (HSPCs) from chemotherapy toxicity while preserving antitumor efficacy. In a subcutaneous tumor model, MSCGlg1 delivers interleukin-7 (IL-7), restoring immune competence without promoting tumor proliferation. This strategy establishes a versatile framework for targeted immunomodulation to treat cancer as a systemic immune disease.

The cytokine thrombopoietin (THPO) promotes both self-renewal and cell cycle quiescence of adult bone marrow (BM) hematopoietic stem cells (HSCs). It remains unclear how THPO differentially regulates HSC expansion versus quiescence and whether it influences the broader BM hematopoietic hierarchy, particularly multipotent progenitor cells (MPPs), which express MPL, the receptor for THPO. Adult constitutional Thpo-knockout mice exhibited significant decrease in both HSC and MPP numbers, yet single-cell RNA sequence-based genetic changes were prominent only in HSCs. Induced deletion of Thpo in adult mice showed that THPO primarily maintains adult BM HSC numbers by inhibiting apoptosis. Thpo deficiency-driven reduction of adult BM MPPs was attributed to the shortage of expansion and differentiation of HSCs during neonatal BM hematopoiesis. Drug-mediated enhancement of THPO signaling in neonatal Thpo-deficient mice rescued adult BM progenitor cell numbers but not HSC apoptosis. THPO function is thus limited during adult hematopoiesis but is critical during neonatal development to establish a structured HSC and MPP hierarchy.

cerebral autosomal dominant arteriopathy with subcortical infarcts and leukoencephalopathy (CADASIL) is a hereditary brain small vessel disease caused by pathogenic variants in the NOTCH3 gene, leading to NOTCH3 protein accumulation and degeneration of vascular smooth muscle cells (VSMCs). Here, we developed a CADASIL 3D Vessel-on-Chip model using either primary brain VSMCs or human induced pluripotent stem cell (hiPSC)-derived VSMCs from CADASIL patients and isogenic controls. In 3D co-culture with hiPSC-derived endothelial cells, both primary and hiPSC-derived CADASIL VSMCs exhibited disease-relevant morphological abnormalities, increased NOTCH3 and contractile protein levels, and altered intracellular Ca2+ dynamics that were not observed under conventional 2D culture. PDGFRβ, a downstream NOTCH3 target, was upregulated and correlated with NOTCH3 protein levels in both 3D models and CADASIL patient brain tissue. Pharmacological inhibition of NOTCH3 cleavage reduced NOTCH3 protein levels and rescued CADASIL VSMC phenotypic abnormalities. In conclusion, this 3D Vessel-on-Chip model robustly shows CADASIL pathology-relevant readouts and provides a platform for mechanistic studies and therapeutic testing.

Neurodegenerative diseases are characterized by progressive neuronal dysfunction and loss. Microglia, the brain's resident macrophages, are key contributors to disease pathogenesis, with many genetic risk variants enriched in microglia-specific genes. While rodent models have provided valuable insights, human induced pluripotent stem cell (iPSC) and embryonic stem cell (ESC) technologies now enable the generation of human microglia-like cells, offering a physiologically relevant platform to study human microglial biology. This review discusses the developmental origins and functions of microglia, current differentiation approaches, and how these models help elucidate disease-relevant phenotypes and molecular mechanisms in neurodegeneration.

Neural crest stem cells (NCSCs), capable of differentiating into neurons and Schwann cells, are essential for peripheral nerve regeneration. This study investigates the role of endogenous NCSC-like cells in mechano-electrical stimulation (MES)-enhanced peripheral nerve repair. In a critical-sized nerve injury model, MES leads to complete nerve reconnection, accompanied by a significant increase in NCSC-like cells at the injury sites. In vitro, MES promotes the simultaneous differentiation of NCSC-like cells into neurons and Schwann cells, with elevated neuregulin 1 (NRG1) expression, a key factor in Schwann cell development. Mechanistically, MES activates BMP/Smad signaling, driving neuronal differentiation and subsequent NRG1 secretion, which in turn promotes Schwann cell maturation through the ErBB/NFAT pathway. These findings demonstrate that MES enhances peripheral nerve regeneration by activating and directing stem cell differentiation, supporting a novel therapeutic approach that utilizes physical stimulation for stem cell modulation for nerve repair.

Although neoadjuvant chemotherapy (NAC) has shown efficacy in reducing tumor burden in colorectal cancer (CRC), its impact on long-term patient outcomes remains limited. Here, we identify quiescent persister tumor cells (PTCs) as a critical determinant of therapeutic failure. Elevated PTC abundance correlates with poor long-term prognosis, even in patients exhibiting an initial response to treatment. Quiescent PTCs possess aggressive stem-like traits and orchestrate an immunosuppressive microenvironment characterized by CD96+CD8+ T cell infiltration in an orthotopic CRC mouse model. CD96 depletion diverts CD8+ T cells from an exhaustion trajectory and promotes memory-like phenotypes through enhanced mitochondrial function. Consistently, anti-CD96 therapy effectively eliminates PTCs in preclinical models. We also engineered epithelial cell adhesion molecule (EpCAM)-targeted human chimeric antigen receptor (CAR)-T cells deficient in CD96 expression, which robustly target PTCs and demonstrate remarkable therapeutic potential against CRC. Overall, our study uncovers CD96 as a previously unrecognized axis of vulnerability within the PTC-driven microenvironment, offering a promising avenue to enhance CRC therapeutic outcomes.

Nasopharyngeal Carcinoma (NPC) tumor stem cells play a crucial role in the occurrence and development of nasopharyngeal carcinoma, but the identity and role of Cancer Stem Cell (CSC) subpopulations that drive metastasis remain unclear. This study explores the mechanisms of NPC stem cell subpopulations (CXCR4+) in the development of NPC, providing a reference for therapeutic targets for NPC.

The regulation of multiple biological reactions in stem cells is linked with the functions of different ion channels, and their modulation is considered as an effective approach to control stem cell behavior. Human endometrial-derived mesenchymal stem/stromal cells (eMSCs) are promising candidates for the use in regenerative medicine due to high availability and minimal ethical issues. The cultivation of eMSCs as 3D spheroids was shown to improve its properties for stem cell therapies. Dramatic changes in cell organization occur during 2D-to-spheroid transition, and this potentially affects eMSC signaling pathways including ion channel-dependent reactions. Previously we have identified BK potassium channels in the 2D culture of eMSCs but their impact to cellular reactions remain unidentified. Here, we aimed to determine and compare the role of BK channel activity in the control of 2D eMSC migration and 3D spheroid reactivation. We observed significant decrease of eMSC spheroid spreading rates in the presence of specific BK channel blockers while no effect of BK inhibitors on 2D eMSC migration was detected. Our results show the 2D-to-3D transition resulted in the inclusion of BK channels to the pathways controlling eMSC migration during spheroid reactivation. Our data demonstrates the particular importance to identify the similarities and differences between key ion channel-controlled signaling pathways and physiological reactions in 2D and 3D cell cultures.