Cell Journal最新文献

Objective: Prenatal prediction of CD34⁺ adequacy supports cord-blood banking by reducing expenses on low-yield units and reserving capacity for clinically promising grafts. We have developed and evaluated a prenatal machine learning model that predicts whether an umbilical cord blood (UCB) unit will meet a clinically supported adequacy threshold (≥1.5×10⁵ CD34⁺ cells/kg recipient) before collection for single-unit grafts.

Materials and methods: In this retrospective study, we analysed 126,406 records from the Royan Stem Cell Technology Company (RSCT; Tehran, Iran), which included routinely available maternal, neonatal, and family-history variables. A pipeline of imputation (IterativeImputer numeric; SimpleImputer+OrdinalEncoder categorical), feature selection (Extra Trees), and hyperparameter tuning using Bayesian optimisation with model training/evaluation was performed within cross-validation folds. Decision Tree (DT), K-Nearest Neighbours (KNN), Random Forest (RF), Support Vector Machine (SVM), and Multilayer Perceptron (MLP) classifiers were tuned via Bayesian optimisation. Models were ranked by the Technique for Order Preference by Similarity to Ideal Solution (TOPSIS). Majority voting (MV) ensembles were constructed from the top-k models. Model interpretability used SHapley Additive exPlanations (SHAP).

Results: The MV (top-4: RF, KNN, DT, MLP) ensemble achieved an area under the receiver operating characteristic curve (ROC-AUC)=0.808 and an area under the precision-recall curve (PR-AUC)=0.744 on the held-out test set, with an accuracy=0.757, precision=0.726, recall/sensitivity=0.804, F1=0.762, specificity=0.716, and Brier score=0.181. SHAP highlighted history of hepatitis C, birth place, hyperthyroidism, history of anaemia, oral fungus, and rheumatism among the most influential features.

Conclusion: Prenatal prediction of UCB CD34⁺ adequacy using an interpretable MV ensemble is feasible and accurate to support pre-collection triage and can potentially improve banking efficiency. The resultant model offers a non-invasive tool to enhance the efficiency of cord blood banking by prioritising units with higher transplantation potential.

Objective: This study intends to determine whether exercise-based interventions improve haemoglobin A1c (HbA1c) levels in adults with type 1 diabetes (T1D) and examines how different types of exercise influence this association.

Materials and methods: A search of the appropriate electronic databases revealed 18 eligible studies from initial 685 records. The effect of exercise training on HbA1c was examined in two parts: i. Exercise training intervention compared to a T1D control group (between-group analysis) and ii. Pre/post comparison of exercise training intervention on T1D patients (within-group analysis).

Results: The between-group results showed that exercise training had a statistically significant reduction in HbA1c [mean difference (MD), confidence interval (CI)]: -0.39 (-0.64, -0.15), P=0.002, I²=0%). Subgroup analysis based on the types of exercise training showed a statistically significant effect of high-intensity interval training (HIIT) on HbA1c levels in both the within-group analyses [MD: -0.24 (-0.45, -0.02), P=0.03, I²=0%] and between-group analyses [MD: -0.39 (-0.64, -0.15), P=0.002, I²=0%].

Conclusion: The meta-analysis and meta-regression results suggest that HIIT may be a supportive intervention in T1D patients; however, there was insufficient evidence to confirm these findings. Data from more randomised controlled trials (RCTs) are needed to compare the effect of different types of exercise on HbA1c levels in T1D patients [registration: PROSPERO (CRD42023414580)].

Objective: The PI3K/Akt signaling pathway plays a central role in regulating cell growth, survival, and metabolism, and its dysregulation is a hallmark of many cancers. The PIK3CA gene, which encodes the alpha catalytic subunit of PI3K, is altered in approximately 30% of breast cancers. Among its mutations, c.3140A>G (p.His1047Arg) in the kinase domain is the most prevalent, producing a constitutively active enzyme with oncogenic potential. Here, we engineered a population of MCF7 cells carrying the PIK3CA c.3140A>G mutation using CRISPR-Cas9 with precise single-nucleotide editing, and evaluated its impact on cellular characteristics.

Materials and methods: In this experimental study, nearly homogeneous populations of PIK3CA H1047R mutant MCF7 cells were generated using CRISPR-Cas9-mediated genome editing followed by hierarchical single-cell isolation. Editing efficiency was validated through allele-specific polymerase chain reaction (PCR) and multiple rounds of Sanger sequencing. Cell cycle distribution and proliferation were analyzed using flow cytometry and cell count assays, respectively. Gene expression changes were assessed by quantitative real-time PCR to evaluate the mutation's impact on cell cycle-related genes.

Results: Tracking of insertions, deletions, and recombination events (TIDER) analysis showed approximately 60% homology-directed repair (HDR) efficiency in the edited population. Flow cytometry revealed a 5% increase in the G2/M cell population in the edited clone compared with unedited controls (P<0.001). Proliferation assays demonstrated significantly accelerated growth (1.30 fold) under low fetal bovine serum (FBS) conditions (P=0.029). Quantitative real-time PCR confirmed upregulation of cell cycle-promoting genes, with CCND1 and MYC expression increasing by 1.62-fold (P<0.001) and 1.23-fold (P<0.001), respectively, relative to controls.

Conclusion: The genetically edited cell lines represent robust and well-defined experimental models that enable direct assessment of the functional consequences of oncogenic driver mutations on cellular behavior and signaling pathways. Our findings demonstrate that targeted genetic alterations induce measurable changes in proliferation, cell-cycle regulation, and gene expression, thereby providing mechanistic insight into tumorigenesis and the specific contribution of driver mutations to cancer-related cellular phenotypes.

Objective: Diabetes is a metabolic disorder with two common categories of complications: microvascular and macrovascular. Cardiovascular diseases (CVDs), which are linked to inflammation in the blood vessels, are the most important complications of diabetes. G protein-coupled receptors (GPCRs) play a crucial role in initiating these inflammatory responses. The aim of this study is to identify GPCRs that may respond to high blood glucose in monocyte cells and have an effect on regulating vascular inflammation.

Materials and methods: In this in-silico and experimental study, first, RNA sequencing (RNAseq) data from monocytes from the blood of diabetic patients were analyzed to find the GPCR most responsive to high blood sugar. Next, the expression correlation of the candidate GPCR with inflammatory genes was calculated, and expression changes in THP-1 cells under hyperglycemia were evaluated via quantitative polymerase chain reaction (qPCR). Finally, the effect of the candidate GPCR on down-regulation of NF-kB mRNA expression, generation of reactive oxygen species (ROS), and caspase activity was evaluated.

Results: The RNAseq analysis showed that expression of Calcitonin receptor-like (CALCRL), one of the GPCR candidates, increased in the monocytes of patients with diabetes compared to healthy individuals. The qPCR result also indicated that expression of the CALCRL gene increased in THP-1 cells treated with 25 mM D-glucose. Correlation analyses demonstrated that increased expression of CALCRL is associated with inflammatory pathways, cytokine production and secretion, and the cell migration process. Finally, our data indicated that CALCRL downregulation decreased inflammation related to hyperglycemia, including NF-kB mRNA expression, ROS production, and caspase 3/7 activity.

Conclusion: Hyperglycemia increased the expression of CALCRL GPCR in the monocytes of diabetic patients. The CALCRL receptor may play a key role in regulating vascular inflammation in hyperglycemic conditions. Targeting CALCRL may control inflammation in blood cells.

Objective: Aggressive cancer cells exhibit many phenotypic and functional similarities to embryonic stem cells (ESCs), including characteristics such as self-renewal, differentiation flexibility, and specific gene expression signatures. While the ESC microenvironment precisely regulates proliferation to balance self-renewal and differentiation, it remains unclear whether tumor cells can respond to these regulatory signals to suppress their tumorigenic phenotype. Extracellular vesicles (EVs) released by human ESCs (hESCs) are known to exert paracrine effects, making them prime candidates for carrying these regulatory signals. The aim of this study was to determine if hESC-EVs suppress the tumorigenic phenotype of aggressive cancer cells by affecting their proliferative, invasive, and stem cell properties.

Materials and methods: In this experimental study, EVs were isolated from hESC-conditioned medium (hESC-CM) and thoroughly characterized. Aggressive breast and melanoma cancer cell lines were treated with the isolated hESCderived EVs (hESC-EVs). The resulting effects on cell growth (proliferation), invasion, stem cell properties (stemness), and overall tumorigenesis were evaluated in vitro and in vivo. Expression levels of key pluripotency markers were also assessed in the treated cancer cells.

Results: The treatment of the aggressive cancer cell lines with hESC-EVs resulted in a significant reduction in proliferation, invasion, stemness, and tumorigenesis across both in vitro and in vivo models. Mechanistically, exposure to hESC-EVs led to the clear downregulation of pluripotency markers within the recipient cancer cells.

Conclusion: These findings suggest that hESC-EVs transmit powerful regulatory signals capable of inducing the loss of stem cell characteristics in aggressive cancer cells. This mechanism offers a potential pathway for reprogramming highly malignant tumor cells toward a more differentiated and less aggressive phenotype, suggesting a novel strategy for anti-cancer therapy.

Objective: Selecting the most competent sperm is crucial for improving the success of intracytoplasmic sperm injection (ICSI). Chemotaxis-based sperm selection may facilitate the selection of spermatozoa with superior motility and fertilization capability. This study aimed to evaluate sperm selection based on chemotaxis induced by microvesicles (MVs) derived from follicular fluid (FF) and cumulus cells (CCs) using a simple microfluidic device.

Materials and methods: In this experimental study, FF and CCs were collected during ovum pick-up from 10 women undergoing ICSI. MVs from both FF and CCs were isolated using the Exocib kit and characterized for particle size, polydispersity index, zeta potential, morphology, and protein content. To establish a chemoattractant gradient, MVs from FF and CCs, as well as progesterone, were incorporated into agar plates. Nine experimental groups were studied: FF- derived MVs (FF-MV, 5, 25, and 50 μg/ml), CC-derived MV (CC-MV, 5, 25, and 50 μg/ml), FF (50 μg/ml), progesterone (0.03 μM), and a control (agar without chemoattractant). Sperm selection was performed in a microfluidic system using semen samples from 20 normozoospermic men. Sperm concentration, motility, progressive motility, viability, and DNA integrity were analyzed following chemotaxis-based selection.

Results: The isolated FF-and CC-derived MVs exhibited spherical morphology, a negative zeta potential, and were polydispersed. The progesterone-containing plates showed sustained release. No significant differences were observed between the CC-MV and control groups in sperm parameters (P=0.630). Plates supplemented with 50 μg/mL of FF-MV markedly improved sperm concentration (P=0.002), progressive motility (P=0.010), and DNA integrity (P=0.001) compared to the control group.

Conclusion: FF-MVs, particularly at 50 μg/ml, possess chemoattractant properties and could selectively enrich sperm populations with improved functional parameters. These findings highlight the potential of FF-MVs as a novel tool for high-quality sperm selection in the ICSI procedure.

Objective: Neuroinflammation plays a crucial role in neuropathic pain, of which toll-like receptor 4 (TLR4) is a key mediator. Virtual screening identified Drospirenone as a potential TLR4 inhibitor. This study aims to evaluate the in vitro effects of Drospirenone on TLR4 signalling in U87-MG astrocytoma cells under different inflammatory conditions.

Materials and methods: In this experimental study, three treatment approaches with Drospirenone were employed to model various stages of neuroinflammation: co-incubation with lipopolysaccharides (LPS) to assess preventive effects; treatment after inflammation to evaluate its impact on sustained inflammation; and delayed treatment after LPS removal to investigate its role in reducing persistent inflammation. Expression levels of TLR4, myeloid differentiation primary response 88 (MyD88), nuclear factor-κB (NF-κB) p65, and interleukin-1 beta (IL-1β) were assessed by Western blotting, while nitric oxide (NO) secretion was measured by ELISA.

Results: The potential ability of Drospirenone to inhibit early-stage inflammation was shown by significant reductions in TLR4, MyD88, NF-κBp65, IL-1β, and NO. Treatment after inflammation showed that Drospirenone reduced NO secretion but did not significantly affect TLR4 and other inflammatory markers, which could indicate its potential efficacy in controlling sustained inflammation. In the delayed treatment approach, inflammation persisted after LPS removal, and Drospirenone did not return the inflammatory state to baseline.

Conclusion: Drospirenone may exhibit potential as a prophylactic agent in vitro during the early phases of neuroinflammation, though its efficacy appears limited in models of chronic or prolonged inflammation. These preliminary findings require in vivo validation, and future studies could explore possible synergistic effects with other treatments or alternative dosing strategies for neuropathic pain management.

Objective: Human embryonic stem cells (hESCs) have the capacity to differentiate into all cell lineages, making them invaluable for cell-based therapies. However, the ability of individual hESC lines to generate the three embryonic germ layers varies. Selecting a cell line with a high propensity for definitive endoderm (DE) differentiation is crucial for producing pancreatic progenitors, hepatocytes, and other endodermal derivatives. Direct and spontaneous differentiation are two commonly used methods to assess hESC differentiation potential; both are complex and time-consuming. This study evaluates a commercially available differentiation kit as a simple and reproducible screening assay to predict the DE differentiation potential of various hESC lines.

Materials and methods: In this experimental study, three hESC lines obtained from the Royan Institute Stem Cell Bank (Tehran, Iran) were used. Pluripotency was confirmed by evaluating cell morphology, OCT4 protein expression, and alkaline phosphatase staining. Early-stage lineage differentiation was compared between the StemMACS™ Trilineage kit and a previously established DE protocol. Differentiation outcomes were assessed using gene and protein expression analyses, including flow cytometry, quantitative real-time polymerase chain reaction (qRT-PCR), and immunostaining.

Results: The three hESC lines exhibited varying capacities for DE differentiation. Flow cytometry analysis of the surface marker CXCR4 showed differentiation efficiencies of 70% for RH2, compared with 35% and 40% for RH5 and RH6, respectively. The superior efficiency of RH2 was consistent across both differentiation protocols, as evidenced by its strong CXCR4 expression using the in-house method.

Conclusion: hESC lines exhibit variable capacities for differentiation. Commercial kit-based screening provides a simple and reliable method to identify suitable hESC lines for generating endoderm-derived cells.

Objective: Breast cancer remains one of the leading causes of mortality among women worldwide. Approximately 80% of patients receive radiotherapy (RT) as part of their treatment. Numerous efforts have been made to distinguish radiosensitive from radioresistant breast tumors. Long non-coding RNAs (lncRNAs) have increasingly been recognized as crucial mediators of cellular responses to radiation among the wide range of biomarkers studied. This study explored the radiation-induced expression changes of three apoptosis-associated lncRNAs-MEG3, MALAT1, and ANRIL-in breast cancer.

Materials and methods: In this experimental study, two established human breast cancer cell lines, MCF-7 and MDAMB- 231, were cultured in Dulbecco's Modified Eagle Medium (DMEM) and irradiated with X-rays at doses ranging from 0 to 8 Gy. The expression levels of the lncRNAs ANRIL, MALAT1 and MEG3 were quantified by real-time polymerase chain reaction (PCR). Cell viability in irradiated MCF-7 and MDA-MB-231 cells was evaluated using the MTT colorimetric assay.

Results: Following radiation exposure, the expression levels of ANRIL and MEG3 were significantly decreased (P=0.004) in both MCF-7 and MDA-MB-231 cell lines, whereas MALAT1 expression was significantly increased (P=0.006) only in MDA-MB-231 cells. In addition, cell viability in both MCF-7 and MDA-MB-231 lines declined after irradiation.

Conclusion: Our findings indicate that radiation exposure increases MALAT1 expression while decreasing ANRIL and MEG3 levels, associated with reduced cell viability .These results suggest a potential role for these lncRNAs in radiation-induced cell death.

There is a high prevalence of infertility worldwide (17.5% of the adult population), with male factors comprising 40% of cases. Therefore, research is vital for addressing the challenges faced by infertile men. Innovative methods, diagnosis, and treatment are necessary for male infertility, and stem cell science has brought about significant breakthroughs in its treatment. Spermatogonial stem cells (SSCs) play a crucial role in maintaining reproductive function in males, and advancements in their in vitro culture show promising developments. An array of specific growth factors and compounds that include epidermal growth factor (EGF), leukaemia inhibitory factor (LIF), glial cell line-derived neurotrophic factor (GDNF), basic fibroblast growth factor (bFGF), stem cell factor (SCF), vitamin A/retinol, testosterone, and follicle stimulating hormone (FSH) are applied to natural or artificial surfaces to mimic the testicular niche to preserve SSCs viability and induce their differentiation. These media should maintain self-renewal ability, continuously generate daughter spermatogonia, and differentiate into spermatozoa. However, several limitations and challenges exist despite the significant progress in SSC culture techniques. One limitation is the difficulty in fully replicating the intricate in vivo testicular niche. This complex microenvironment that includes the dynamic interplay of somatic cells, hormones, growth factors, and external matrix components is crucial for supporting SSC self-renewal and differentiation. Currently, in vitro systems do not adequately replicate this microenvironment, resulting in suboptimal SSC maintenance and inconsistent differentiation outcomes. This might be a key challenge for achieving complete and functional spermatogenesis in vitro. We review the foundational concepts and recent breakthroughs in in vitro spermatogenesis, focusing particularly on studies in human systems from the past twenty years. As a conclusion from this review article, developing a defined culture system that accurately mimics the in vivo niche is crucial for maintaining the functional properties of spermatogonial stem cells. The establishment of such a reliable platform is an essential prerequisite for translating in vitro spermatogenesis into clinical applications for treating infertility.