Naphthoquinones mediate differentiation of human umbilical cord derived mesenchymal stem cells into insulin producing cells through regulation of Wnt and BMP pathways

Urine-Derived Stem Cells: Challenges in Isolation, Biological Identity, and Therapeutic Potential in CKD-Associated Fibrosis

Urine-derived stem cells (UDSCs) have emerged as a promising cell source for regenerative medicine due to their non-invasive procurement, high proliferative capacity, and potential relevance to kidney-specific repair. Unlike conventional mesenchymal stem cells (MSCs) obtained from bone marrow or adipose tissue, UDSCs originate from multiple regions of the urinary tract and exhibit a unique biological profile that combines MSC characteristics with features of renal progenitor populations. This review provides a comprehensive overview of the current understanding of UDSC biology, including their origin, isolation strategies, morphology, immunophenotypic characteristics, differentiation potential, and secretory profile. Particular attention is given to the expression of renal lineage-associated markers and pluripotency-related factors that may contribute to their regenerative capacity. The bioactive mediators of UDSCs regulate inflammation, oxidative stress, angiogenesis, and extracellular matrix remodeling, thereby influencing key pathways implicated in chronic kidney disease (CKD)-associated fibrosis. Furthermore, the intrinsic renal progenitor signature of UDSCs may provide advantages in renal homing and tissue-specific repair compared with conventional MSC populations. Despite encouraging preclinical findings, significant challenges remain, including cellular heterogeneity, inconsistent isolation efficiency, lack of standardized characterization criteria, and limited clinical validation. Collectively, current evidence positions UDSCs as a biologically distinct and therapeutically attractive platform for kidney regeneration.

Differentiation potential of human amniotic fluid derived-mesenchymal stem cells into dopaminergic neurons.

Stem cell-based therapies have emerged as promising alternatives for disease modification and cell replacement, aiming to improve the quality of life in patients with Parkinson's disease. Human amniotic fluid mesenchymal stem cells (hAF-MSCs) represent a viable source due to their self-renewal ability, high proliferative capacity, and multipotent differentiation potential. This study aimed to evaluate the in vitro differentiation potential of hAF-MSCs into dopaminergic neurons. Flow cytometry and Alamar blue assays were employed to confirm mesenchymal stem cell characteristics. The hAF-derived cells fulfilled the established MSCs criteria, including fibroblast-like morphology, surface marker expression, and proliferative capacity. Upon induction with dopaminergic differentiation factors, the cells acquired neuron-like morphology. RT-qPCR analysis revealed significantly upregulated expression of neuronal and dopaminergic-specific genes (nestin, β-III-tubulin, and tyrosine hydroxylase), further supported by immunofluorescent staining showing the presence of neuron-related proteins. These findings demonstrate the potential of hAF-MSCs to differentiate into dopaminergic neurons and highlight their promise for future applications in regenerative medicine.

Type 2 diabetes mellitus limits the glucose-responsive β-like functionality but not pancreatic lineage commitment of induced human adipose-derived stem cells.

BACKGROUND Type 2 diabetes mellitus (T2DM) is one of the most common chronic diseases, whose prevalence is increasing worldwide. There is a great demand for an effective cure that attains a normal glucose level. The differentiation of mesenchymal stem cells into insulin-producing cells (IPCs) for pancreatic regeneration represents a promising anti-diabetic therapeutic approach. However, the pathogenic milieu of T2DM may affect their differentiation potential. AIM This study aimed to investigate the impact of the T2DM milieu on the pancreatic differentiation potential of human adipose-derived stem cells (ASCs) in vitro. METHODS We isolated and cultured ASCs from T2DM (dASCs) and non-diabetic (nASCs) subjects, then we differentiated the cells into IPCs. We evaluated the differentiation potential using dithizone staining, immunofluorescence staining, and gene expression analysis. We examined the functionality of generated cells via glucose challenge assay. RESULTS Our results revealed that both dASCS and nASCs could be committed to an early endocrine-like phenotype with comparable potential. However, the glucose-stimulated insulin secretion of dASCs-derived IPCs was significantly less than that derived from nASCs. CONCLUSION T2DM does not affect the pancreatic commitment potential of dASCs. However, ASCs-derived from diabetic patients displayed a reduced capacity to acquire glucose-responsive endocrine characteristics following pancreatic differentiation, exhibiting a limited β-like functionality.

Cell-free MSC secretome therapy ameliorates diabetic testicular injury via antioxidant mechanisms

Electrical Stimulation of Human Adipose Tissue-Derived Mesenchymal Stem Cells and Schwann Cells for Regulating Extracellular Vesicle Biogenesis and Inflammation

Peripheral neuropathy (PN) is a debilitating condition characterized by chronic pain, numbness, and motor dysfunction, with limited treatment options. Ischemic stroke can cause central neuropathy, which may also induce PN. Human mesenchymal stem cells (hMSCs) have shown promise in therapeutic applications, but limitations in cell viability, immune response, and efficacy persist. Extracellular vesicles (EVs), which facilitate cell-free intercellular communication, offer a promising alternative for nerve regeneration. Electrical stimulation (ES) has emerged as a method to enhance EV secretion, and this study investigates its potential for promoting EV production from human adipose tissue-derived mesenchymal stem cells (hASCs) and human Schwann cells (hSCs). In this study, hASCs, hSCs, and lipopolysaccharide (LPS)-induced inflamed hSCs were subjected to one hour of low-frequency direct current (DC) electrical stimulation (100 mV/mL) for 7 days. EVs were isolated using differential ultracentrifugation and characterized through nanoparticle tracking analysis (NTA). Gene expression was analyzed via qRT-PCR to evaluate markers associated with EV biogenesis as well as pro- and anti-inflammatory cytokines. Our results demonstrate that ES significantly increases EV secretion from both hASCs and hSCs, with a notable upregulation of genes involved in both the endosomal sorting complex required for transport (ESCRT)-dependent and ESCRT-independent pathways of EV biogenesis. Additionally, ES modulates inflammation-related markers, promoting anti-inflammatory gene expression and reducing pro-inflammatory gene levels. Notably, LPS-induced hSCs exhibited a phenotype shift from myelinating to non-myelinating cells, producing EVs capable of modulating the inflammatory microenvironment. However, prolonged exposure to ES led to a decrease in EV secretion and changes in EV size distribution, suggesting potential cellular adaptation or membrane stress. This study highlights the potential of ES as a scalable, cell-free strategy to enhance EV production, offering new insights into its therapeutic applications for peripheral neuropathy and nerve regeneration.

Effects of hypoxia on the biological characteristics of urine-derived stem cells

Introduction Urine-derived stem cells (USCs) are a promising stem-cell source because they can be obtained non-invasively and are readily available. However, their biological characteristics under hypoxic conditions remain insufficiently characterized. This study evaluated the effects of hypoxia on USCs in vitro. Methods USCs isolated from six healthy male donors were cultured under normoxic (21% O2) or hypoxic (3% O2) conditions. Colony formation, viability, proliferation, cell cycle, apoptosis, migration, surface markers, stemness-related gene expression, multilineage differentiation, cytokine secretion, and transcriptomic changes were assessed. Results Hypoxia enhanced colony formation, proliferation, migration, stemness-related gene expression, and osteogenic, adipogenic, and chondrogenic differentiation, while maintaining cell viability and not significantly affecting apoptosis or surface marker expression. VEGF secretion increased, whereas other assessed cytokines showed no significant differences. RNA sequencing identified 1,267 differentially expressed genes, including 748 upregulated and 519 downregulated genes. Hypoxia upregulated oxygen-sensing, metabolic, and HIF-1 pathways and downregulated Hippo signaling. Discussion Hypoxia at 3% O2 enhanced multiple biological characteristics of USCs without compromising viability. These findings provide a foundation for further investigation of hypoxia-conditioned USCs in regenerative medicine.