Intrauterine adhesion (IUA), a prevalent cause of female infertility and recurrent pregnancy loss, is characterized by endometrial trauma and progressive fibrosis. Current treatment modalities, including hysteroscopic adhesiolysis and hormone decidual multipotent mesenchymal stromal cells (DMSCs), a unique subset of stromal cells derived from the endometrium, exhibit strong multipotent differentiation capabilities, immunomodulatory properties, and low immunogenicity. These features enable DMSCs to facilitate endometrial regeneration, restore intrauterine immune homeostasis, and attenuate fibrosis, offering a compelling therapeutic strategy for IUA. Recent preclinical studies have demonstrated promising regenerative outcomes, yet the clinical application of DMSCs remains constrained by challenges such as limited cell availability, variability in therapeutic efficacy, and concerns regarding long-term safety. This review provides a comprehensive overview of the current progress in DMSC-based therapy for IUA, highlights its mechanistic advantages, and discusses critical obstacles and future directions for successful clinical translation.

Human induced pluripotent stem cells (hiPSC) are an invaluable resource for investigating the molecular mechanisms regulating cell fate specification during brain development. However, most directed differentiation methods exhibit significant cell fate heterogeneity and require several months to become functional. To address this challenge, we developed a green fluorescent protein (GFP) reporter system in hiPSC by targeting the genomic locus of Forebrain Enriched Zinc Finger 2 (), which encodes a transcription factor essential for the fate specification of sub-cerebral projection neurons (SCPN) during forebrain development. Using this FEZF2-GFP reporter hiPSC line, we optimized a directed differentiation protocol to rapidly and efficiently generate pallial progenitors and glutamatergic neuronal subgroups after 3 weeks. Through fluorescence activated cell sorting for both GFP and CD200, isolated post-mitotic SCPN immediately displayed electrophysiological properties and formed glutamatergic synapses within 4 additional weeks of cell culture. Co-culture with hiPSC-derived spinal motor neurons further enhanced these electrophysiological characteristics, improved viability, and increased synapse formation in SCPN. This study presents a streamlined and effective strategy to generate, isolate, and characterize human motor neuron circuits, providing insights into the molecular determinants regulating synaptogenesis and functional maturation.

Hematopoietic stem cell (HSC) transplantation improves stroke outcomes. The mechanism of HSC transplantation involves delivering metabolites, such as glucose, to injured cerebral endothelial cells via gap junctions. To mimic HSC function, we prepared glucose-encapsulated liposomes functionalized with sialyl Lewis X on their surfaces and evaluated their therapeutic effects in a murine stroke model. As a result, liposomes with sialyl Lewis X accumulated in both the poststroke and contralateral cortices, whereas those without sialyl Lewis X showed no accumulation. Administration of glucose-encapsulated liposomes with sialyl Lewis X improved stroke outcomes and enhanced cerebral blood flow. Our findings indicate that liposome therapy could serve as a promising alternative to stem cell therapy for stroke.

Vitamins are well known for their essential role in maintaining organismal homeostasis by directly influencing the function of a variety of cells. Among the tissues crucial for sustaining life is the bone marrow, where the highly organized and demanding daily production of billions of new blood cells occurs through a process known as hematopoiesis. At the apex of the hematopoietic hierarchy lie hematopoietic stem cells (HSCs), undifferentiated cells with multilineage potential that maintain themselves through extensive networks of cell-intrinsic and cell-extrinsic interactions. Despite considerable efforts to unravel these regulatory networks, the ways in which most vitamins influence the delicate balance of blood homeostasis remain largely unexplored. In this review, we summarize the current body of evidence linking vitamins to HSC metabolism and progeny, with a particular focus on preclinical findings that highlight the role of vitamins in maintaining hematopoietic tissue function. A deeper understanding of the contributions of vitamins to blood cell production could pave the way for innovative therapeutic approaches against hematological diseases.

Cells with renal progenitor cells characteristics are shed in urine. This study aimed to investigate whether these SIX2-positive urine-derived renal progenitor cells (UdRPC) have therapeutic potential in treating or managing acute to chronic kidney injuries, which are increasing worldwide and currently affect one in ten people. Human UdRPC were obtained from a 35-year-old woman, expanded, and characterized in vitro before being transplanted unilaterally under the renal capsule of mouse kidneys that had undergone ischemia reperfusion injury (IRI). The blood sera of the mice were examined for kidney injury markers such as blood-urea-nitrogen and creatinine, and proteome changes over the 21-day study period using proteome arrays and bioinformatic methods. At the end of the study period, connective tissue deposition in the kidneys was examined histologically through Sirius Red staining, and the expression of fibrosis- and inflammation-associated genes was determined by reverse transcription quantitative polymerase chain reaction. The blood serum analysis revealed that the transplanted human UdRPC transiently influenced the secretome in mice. Furthermore, the transplanted cells improved renal fibrosis resulting from IRI, indicating therapeutic relevance. Deposition of extracellular matrix proteins and the expression of fibrosis-associated genes, such as connective tissue growth factor as well as collagen 1α2 and 3α1 chains, significantly decreased in the injured kidney after UdRPC transplantation. The expression of genes linked to inflammation and chronic kidney disease, such as monocyte chemoattractant protein 1 and intercellular adhesion molecule 1, was also reduced in mouse kidneys in the presence of UdRPC. K-means cluster analysis of the serum proteome from mice with IRI and transplanted UdRPC at different time points revealed a transient increase in immunomodulatory, antifibrotic, and angiogenic factors that could have triggered these positive effects on acute to chronic mouse kidney disease. Transplanted human UdRPC alleviated the kidney injury severity through ameliorating fibrosis; however, they could not restore complete kidney function within 21 days.

Mesenchymal stromal cells (MSCs) are currently used in clinical practice as a therapeutic agent for immunomodulation and tissue repair. They are found in all supporting tissues, including perinatal tissues such as umbilical cord and amniotic membranes (amnion and chorion). Perinatal tissues have attracted interest due to their availability, minimal ethical and legal concerns, and high banking potential for allogeneic applications. Many studies have compared the efficacy of MSCs from different sources, without reaching a consensus on the most effective to use in a given clinical situation. This study compared the transcriptomic signatures of MSCs derived from adult bone marrow (BM-MSCs)-the reference source most widely used in clinical trials-with those of perinatal MSCs (P-MSCs). Our data were analyzed jointly with three independent transcriptome datasets. Unsupervised principal component analysis revealed a major stratification according to tissue origin, accounting for 16.6% of the total transcriptomic variance, without any detectable bias from batch effects or cell culture procedures. Supervised differential expressed gene analysis between BM and perinatal samples revealed 819 genes presenting differential expression. Gene Set Enrichment Analysis highlighted that adult BM-MSCs are implicated in adipogenesis and osteoblast differentiation, whereas P-MSCs upregulated gene sets implicated in cell cycle regulation, functions classically described in the literature. Among the different sources of variability, we showed that perinatal tissues have a strongly distinct transcriptional signature compared with adult BM, independent of the production center or the culture conditions used. The in-depth study of transcript profiles therefore seems to remain a valuable and robust characterization tool for cell therapy banking.

Cannabis use during pregnancy is increasing. In rodent models of delta-9 tetrahydrocannabinol (Δ9-THC) exposure during pregnancy, placental pathology, including compromised labyrinth development, is reported. Cannabinoid receptor 1 (CB1/) is the primary mediator of Δ9-THC action, with its expression reportedly limited to the placental junctional zone in the rodent. Given a Δ9-THC-induced labyrinth-specific pathology, we predicted more diverse expression. This study aimed to elucidate the spatiotemporal expression of CB1/ in the rodent and assess whether it mediates Δ9-THC-induced alterations in trophoblast differentiation. Results revealed CB1 expression in all maternal blood-facing trophoblast cells. Furthermore, Δ9-THC exposure (at levels matching those reported in maternal serum) had a more significant effect on the expression of markers associated with differentiating trophoblast cells than on proliferating trophoblast stem (TS) cells. Δ9-THC impacted mouse (m) TS cell differentiation in a CB1-dependent manner, reducing the expression of syncytiotrophoblast (SynT) markers, driving differentiation along the junctional zone/trophoblast giant cell pathway. mTS cells without CB1 (mTS) did not express markers of SynT cells or the differentiated junctional zone cell types. However, at a higher than physiologically relevant concentration, Δ9-THC (15 μM) induced (SynT) expression in mTS cells. This study reveals a mechanism by which Δ9-THC may impact placental growth.

Metastasis is the primary cause of death in advanced/recurrent cancer patients. Cancer metastatic capability depends not only on cancer cells but also on the cancer microenvironment, particularly cancer-associated fibroblasts (CAFs), a highly heterogeneous population. Our prior work identified a POSTN-secreting CAFs subpopulation linked to gastric cancer (GC) invasion and poor survival. The Cancer Genome Atlas analysis in GC showed POSTN association with epithelial-mesenchymal transition and extracellular matrix degradation pathways. In vitro, GC exosomes induced adipose-derived mesenchymal stem cells (MSCs) into POSTN-expressing CAFs. Lentiviral POSTN overexpression in CAFs enhanced GC cell migration/invasion, while knockdown had the opposite effect. These results were validated in a nude mouse GC model. As POSTN is an integrin ligand, POSTN-positive CAFs (POSTN+ CAFs) activated integrin downstream AKT signaling. AKT inhibition significantly diminished the pro-migratory/invasive effect of POSTN-overexpressing CAFs. In summary, POSTN+ CAFs promote GC invasion via AKT pathway activation.

This study investigates the effects of inflammation and orthodontic forces on the osteogenic differentiation of human dental follicle stem cells (hDFSCs) and their role in periodontal regeneration during orthodontic tooth movement (OTM) in periodontitis patients. Human DFSCs were exposed to interleukin-1β (IL-1β) and orthodontic compressive force (OCF). Western blot and quantitative real-time polymerase chain reaction (qRT-PCR) assessed the expression of a disintegrin and metalloproteinase with thrombospondin motifs 2 (ADAMTS2), osteogenic markers, and ERK signaling components. ADAMTS2 overexpression (OE) plasmids and ERK activators were applied to explore the ADAMTS2/ERK signaling axis. A mouse periodontitis model with OTM was developed for in vivo evaluation. IL-1β and OCF downregulated ALP, RUNX2, and ERK pathway-related proteins and decreased ADAMTS2 expression. OE of ADAMTS2 significantly enhanced ERK phosphorylation ( < 0.05), promoting osteogenic differentiation of hDFSCs. ERK pathway activation with C16-PAF partially reversed the suppression of osteogenic differentiation induced by IL-1β and OCF. ADAMTS2/ERK axis components and osteogenic markers were reduced on the compressed side of periodontal tissues in the PD+OTM group ( < 0.05). These findings suggest that ADAMTS2 regulates the osteogenic differentiation of hDFSCs via the ERK signaling pathway under IL-1β and OCF stimulation, highlighting its potential as a therapeutic target for alveolar bone regeneration during OTM in periodontitis patients.

The cementum is a mineralized tissue that covers the tooth root and is necessary for anchoring periodontal ligament fibers to the teeth. During root development, dental follicle (DF) cells differentiate into cementoblasts, periodontal ligament fibroblasts, and osteoblasts to form the cementum, periodontal ligament, and bone, respectively. However, the mechanisms underlying these cell fate decisions remain unclear. Using scanning electron microscopy, we observed a basement membrane (BM)-like structure between the forming root dentin on the periodontal ligament side and the Hertwig's epithelial root sheath (HERS), as well as similar BM structures on the dentin surface after HERS removal. We hypothesized that these HERS-derived BM structures are involved in the differentiation of DF cells into cementoblasts and investigated the effect of basement membrane components (BMCs) on the differentiation of DF cells. Using a cell line (MDF) derived from DF cells of the incisors, we studied adhesion to hydroxyapatite. The undifferentiated MDF cells showed no adhesion. However, when coated with the BMC secreted by the HERS cell line (HERS02T), established from the first lower molars of tdTomato mice, MDF cells demonstrated increased initial adhesion, alkaline phosphatase activity, and calcification ability on hydroxyapatite. Additionally, we performed a comprehensive proteomic analysis of BMC secreted by HERS02T cells and-based on total spectral counts-identified three major proteins: laminin-332, tenascin-C, and periostin. These protein components were coated onto apatite-coated dishes and tested for the induction of MDF differentiation. The results showed that the coating significantly upregulated the expression of the cementoblast differentiation markers and . These results suggested that HERS may detach after BM formation on dentin surfaces. Subsequently, DF differentiate into cementoblasts using the BM when they migrate to the dentin surface.

Epithelial tissues rely on tightly regulated stem cell populations to sustain self-renewal and repair, with fundamental signaling pathways and molecular mechanisms playing conserved roles across species. While mammalian models, particularly mice, have been widely used to study these processes, the cost, complexity, and ethical considerations associated with these models necessitate complementary approaches. The midgut has emerged as a powerful model system for studying epithelial stem cell biology, providing insights into homeostasis, aging, and cancer. The genetic tools, affordability, and rapid experimental timeline of make it an ideal system for investigating fundamental principles of epithelial stem cell regulation. Complementary use of alongside mammalian and advanced models such as organoids has the potential to accelerate discoveries in homeostasis, aging, and cancer biology.

p21 is a cell cycle regulator that has been implicated in regeneration of tissues and in development of certain tumors. p21 inhibition also enhances bone regeneration after injury in p21 mice. To translate these findings to the clinic, we sought an FDA-approved p21 attenuator. UC2288, a derivative of sorafenib, selectively inhibits p21 independently of p53 and induces apoptosis in cancer cells. Given the central role of p21 in mesenchymal stem cell (MSC) proliferation and differentiation, its effects on MSCs merits investigation but remains unknown. Consequently, we hypothesized that UC2288 will improve the osteogenic potential of mesenchymal stem cells by suppressing p21. First, we examined the differential interaction of UC2288 with human bone marrow (BM) MSCs compared with breast cancer cells via viability assays. Increased cell death was observed in cancer cells, particularly at higher concentrations and with longer interaction times, whereas MSCs demonstrated lower cell death. Gene expression assay revealed upregulation of osteogenic genes, though the specific genes overexpressed varied depending on the culture medium. Interestingly, the culture medium also affected p21 expression, where p21 expression was upregulated in DMEM/F12 and downregulated in alpha-MEM as evidenced in gene and protein expression assays. Alizarin Red staining confirmed increased mineralization when UC2288 or UC2288+osteogenic factors were added. These findings indicate that UC2288 promotes osteogenesis in BM-MSCs in a concentration- and time-dependent manner. Further research is needed to optimize conditions for preclinical and clinical translation as an anabolic bone formation therapy.

This study evaluates the clinical efficacy and safety of intravenous allogeneic mesenchymal stem cell (MSC) therapy as an adjunct to standard immunosuppressive treatment in dogs diagnosed with primary immune-mediated hemolytic anemia (IMHA), particularly in cases with inadequate response to conventional immunosuppressive protocols. A total of 157 client-owned dogs with IMHA received MSC therapy at Safari Veterinary Care Center or collaborating clinics. A core cohort of 43 dogs met strict inclusion criteria: confirmed primary IMHA, no significant comorbidities, and receipt of at least two MSC treatments. A retrospective analysis was performed on medical records of treated dogs. Packed cell volume (PCV) trends were used to assess treatment response, with success defined as sustained improvement to ≥30% PCV following MSC therapy. Safety was assessed by reviewing for adverse reactions postinfusion. In the core cohort, 76% (33/43) of dogs achieved successful hematological recovery. Across the broader cohort, clinical improvement was also observed, particularly in dogs with poor initial response to standard therapy. No serious adverse events were recorded following MSC infusion. Allogeneic MSC therapy appears to be a well-tolerated and potentially effective adjunctive treatment for canine IMHA, particularly in refractory cases. These findings support further investigation in controlled clinical trials.

Ensuring the safety of drugs during pregnancy is a critical concern in clinical practice. Animal tests are not always reliable for accurately identifying human teratogens, as evidenced by the infamous thalidomide case. Therefore, there is an urgent need for rapid, dependable, and cost-effective human in vitro assays for potential teratogenic drugs. Here, we utilized human embryoid body (hEB)-based assays, both serum-free and supplemented with pregnant woman serum, along with RNA-sequencing analyses, to demonstrate the effectiveness of hEB transcriptomics as a powerful tool for identifying potential teratogenic drugs, in comparison with other cell systems. Additionally, we subjected our hEB systems to C doses of four nucleoside/nucleotide analogs (adefovir dipivoxil [ADV], entecavir [ETV], lamivudine [LAM], and tenofovir disoproxil fumarate [TDF]), which are used for anti-hepatitis B virus treatment and belong to different pregnancy-related risk categories. Transcriptomics after drug treatments were determined and analyzed for important changes in genes, gene categories, signaling pathways, and cell lineages. TDF and ADV appear to be safer options during the very early stages of embryonic development compared with LAM and ETV. Our findings indicate that EB transcriptomics-based analyses can accurately identify potent teratogens and aid in assessing the potential risk of commonly prescribed drugs during pregnancy.

Sickle cell disease (SCD), affecting approximately 2.1% of Bahrain's population, is a prevalent inherited disorder that necessitates effective treatments and long-term management. This review highlights two innovative gene therapies (Casgevy and Lyfgenia) and compares their efficacy and safety with hematopoietic stem cell transplantation (HSCT)-the only curative option currently available for SCD. While HSCT offers a 90% success rate with suitable donors, its limitations include donor scarcity and toxicity. Gene therapies like Casgevy and Lyfgenia show promising efficacy in reducing SCD complications while bypassing such limitations. In the Kingdom of Bahrain, the Bahrain Oncology Center approved Casgevy in December 2023 and completed its first patient treatment in mid-February 2025, making Bahrain an early adopter. This milestone marks a crucial moment in the history of both SCD and gene therapies and thus warrants exploring the considerations revolving around their implementation. Although these therapies seem to offer hope for patients ineligible for HSCT, their long-term outcomes remain unassessed-further studies with extended follow-up are needed to confirm their safety and durability.

Stem cell therapy employing stem cells from human exfoliated deciduous teeth (SHED) has demonstrated efficacy in treating peripheral nerve injury; however, the precise underlying mechanisms remain largely undefined. In this study, we investigated the effects of SHED on signal transducer and activator of transcription 3 (STAT3), a key mediator of inflammation following sciatic nerve injury (SNI). The left sciatic nerve was transected (cut group), sutured and wrapped with cellulose (suture group), or sutured and enveloped with SHED-soaked cellulose (SHED group). The L4-5 segments of the spinal cord were harvested up to 7 days post-SNI, and tissues were separated into ipsilateral and contralateral regions for molecular and immunohistochemical analyses. In the SHED group, the sciatic functional index showed significant improvement compared with the suture group beginning at 4 weeks postinjury, and tibialis anterior muscle mass was markedly restored at 12 weeks. STAT3 phosphorylation at Tyr (-STAT3) was prominently elevated between 12 and 48 h post-SNI on the ipsilateral side, but not contralaterally. This phosphorylation was localized to motor neurons in the anterior horn and was substantially attenuated by SHED administration between 24 and 48 h postinjury. Moreover, interleukin (IL)-6 expression was significantly reduced at 12 h, while -STAT3 and importin β1 levels were notably decreased between 12 and 24 h. Erk signaling was significantly activated in S100β-positive Schwann cells (SCs) on day 4 at the site of SNI in the SHED group. These results suggest that SHED mitigate neuroinflammation by suppressing IL-6 expression and modulating STAT3 activation, while concurrently enhancing remyelination through Erk signaling activation in SCs at the injury site. Collectively, these findings underscore the therapeutic promise of SHED as a potent and innovative intervention for peripheral nerve avulsion injuries.

The dental pulp not only serves as the tooth's nutritional core but also creates a finely tuned microenvironment that is enriched with blood vessels, nerves, extracellular matrix components, and signaling molecules, all of which guide the fate of resident dental pulp stem cells (DPSCs). Trauma and microbial invasion disrupt this niche, leading to pulpitis and necrosis. Although conventional root canal treatment preserves the tooth's structure by removing infected pulp, it can increase tooth brittleness and impede root development in immature permanent teeth. Harnessing DPSCs' multipotency for pulp regeneration promises to restore the natural pulp-dentin complex in situ. Importantly, DPSCs encounter an inflammatory microenvironment composed of pathogen-associated molecular patterns, a spectrum of pro- and anti-inflammatory cytokines, diverse immune cell phenotypes, and altered matrix signals. While earlier work examined the isolated effects of mediators such as lipopolysaccharide, tumor necrosis factor-alpha, or macrophage-derived exosomes on odontogenic differentiation, this review focuses on how these mediators collectively interact in both synergistic and antagonistic ways within the inflammatory niche. We systematically delineate how these collective stimuli converge on wingless/integrated/beta-catenin, mitogen-activated protein kinase, nuclear factor kappa-B (NF-κB), and bone morphogenetic protein/Sma and Mad related protein pathways to modulate key odontogenic markers (runt-related transcription factor 2, dentin sialophosphoprotein, dentin matrix protein 1, alkaline phosphatase) and mineralization outcomes. By applying a microenvironment-centric lens, we reveal novel targets and strategies to recalibrate inflammation, steer DPSCs toward reparative odontogenesis, and ultimately enhance the efficacy of regenerative endodontic therapies.

Amelogenin has been widely used in clinical practice for periodontal bone regeneration. However, the precise mechanism underlying its osteogenic effects remains incompletely understood. In this study, we hypothesized that amelogenin enhances periodontal bone regeneration by facilitating the migration and homing of bone marrow mesenchymal stem cells (BMMSCs). BMMSCs were used to evaluate the cell migration promoting ability of amelogenin by the Transwell test. Immunofluorescence was performed to assess the beta-catenin nuclear translocation following amelogenin treatment. To investigate amelogenin-induced cell homing in vivo, we established a green fluorescent protein (GFP)-labeled bone marrow transplantation model using BALB/c mice transgenic for GFP. The migratory effects of amelogenin were examined in this model, with Wnt3a, a Wnt/β-catenin pathway activator, serving as a positive control. Subsequently, cell homing and bone regeneration were evaluated through a fluorescence microscope, micro-CT, hematoxylin and eosin (H&E), and Masson staining. In vitro Transwell assays demonstrated that amelogenin significantly enhanced BMMSC migration, with effects comparable with Wnt3a, a canonical Wnt/β-catenin pathway activator. Immunofluorescence analysis revealed pronounced nuclear translocation of β-catenin in BMMSCs following a 24-h amelogenin treatment. Notably, these effects were abolished by a Wnt/β-catenin pathway inhibitor, confirming the pathway's involvement. In GFP-labeled bone marrow-transplanted mice, amelogenin treatment significantly increased GFP cell recruitment to the bone defect site, mirroring the effects of Wnt3a. Micro-CT and histological (H&E) analyses further demonstrated that both amelogenin and Wnt3a accelerated bone regeneration compared with untreated controls. Crucially, this regenerative effect was suppressed upon Wnt/β-catenin pathway inhibition, reinforcing the mechanistic link between amelogenin and β-catenin-mediated osteogenesis. Amelogenin and Wnt3a promoted periodontal bone regeneration both in vitro and in vivo by enhancing BMMSC migration through Wnt/β-catenin signaling activation.

In the original publications of Yamanaka et al. from 2006 to 2007, which were the basis for the Nobel Prize in medicine, murine, and human fibroblasts had been used as the primary cell source for the generation of induced pluripotent stem cells (iPSCs). Over time, four other types of somatic cells have been revealed to be suitable for pluripotency induction, namely blood cells, keratinocytes, urine-derived epithelial cells, and mesenchymal stem cells. Although mature cells have been frequently used for the generation of iPSCs, numerous primary cell types have also been reprogrammed successfully. In this review, we address the current state of research dealing with different sources of human somatic cells used for the generation of iPSCs. Our objective is to provide a comprehensive tabular summary of the sources of somatic cells, organized according to the four main types of tissue (connective tissue, epithelial tissue, muscle tissue, and neural tissue). This overview will serve as a guide for researchers new to the field looking for suitable sources to generate their own iPSCs, for those interested in generating patient-specific iPSCs, or for those seeking further literature on specific cell sources.

The extracellular vesicles (EVs) secreted by mesenchymal stromal cells (MSC-EVs) exhibit immunoregulatory functions dependent on their parent cells. MSC-EVs are promising candidates for treating neuroinflammation in neurological diseases due to their acellular nature and their ability to reach the central nervous system. However, the conditions of MSCs for producing EVs with the highest anti-inflammatory efficacy are still unknown. Therefore, the first objective was to study the characteristics of the EVs produced by MSCs cultured in different conditions. The second objective was to evaluate the anti-inflammatory properties of those EVs in feline stimulated mixed glia. Umbilical cord-derived MSCs were treated with serum-free (SF) media, inflammatory (IF) media, or media supplemented with 5% EV-depleted fetal bovine serum (FBS). The isolated MSC-EVs were characterized by particle size and yield, and their anti-inflammatory ability was evaluated in lipopolysaccharide (LPS) stimulated feline mixed glia. All EV isolates were <160 nm, and the primary mixed glia consisted of microglia, astrocytes, neurons, and endothelial cells. Our results indicate that IF-EVs statistically significantly decreased the production of interleukin 6 (IL-6) and tumor necrosis factor alpha (TNF-α) and downregulated the transcription of the, nuclear factor kappa B p65 subunit in inflammatory mixed glia after 48 hours. In addition, SF- and FBS-EVs significantly reduced the secretion of IL-6 after 48 hours but only SF-EVs achieved a significant effect on inhibiting the expression of p65 at 48 hours. Moreover, messenger RNA (mRNA) levels of inducible nitric oxide synthase (iNOS) were significantly decreased following treatment with SF-EV for 24 hours. This study demonstrates that MSC culture conditions affect the therapeutic potential of the secreted EVs in feline mixed glia.

The study of skeletal muscle disorders in patients with mitochondrial diseases is crucial for gaining insights into disease physiology; however, their molecular mechanisms have not been fully elucidated. We previously established human-induced pluripotent stem (iPS) cells in two patients with the mitochondrial DNA (mtDNA) A3243G mutation and isolated iPS cell clones with either undetectable or high levels of mutations. In the present study, we established skeletal muscle cells from iPS cells with mutation-high and mutation-undetectable clones and comparatively analyzed their mitochondrial functions. Fluorescence immunostaining, fusion index, and qRT-PCR revealed no differences in the morphology, differentiation efficiency, or expression levels of skeletal muscle markers between the mutation-high and mutation-undetectable clones. However, the basal oxygen consumption rate, an indicator of mitochondrial respiration, and adenosine triphosphate (ATP) production were reduced in the mutation-high clones of patients 1 and 2. In addition, the extracellular acidification rate, an indicator of glycolytic activity, was reduced in mutation-high clones of patient 2, who exhibited a more severe clinical phenotype. In the mutation-high clones of both patients, mitochondrial Complex I activity and mtDNA copy number were also reduced, whereas the expression levels of peroxisome proliferator-activated receptor gamma coactivator 1α and glucose transporter type 4 were upregulated, indicating compensation for ATP deficiency. These findings reveal the effects of mitochondrial disorders on energy metabolism in skeletal muscles and provide novel insights into skeletal muscle dysfunction in patients with mitochondrial diseases.