Midlands Innovation Extracellular Vesicles Network (MIEVN) is a joint partnership across the 7 Midlands Innovation Health institutions which aims to:
■ Develop a vibrant multidisciplinary research community, bringing focus to the diverse Extracellular Vesicle (EV) research activity in the Midlands both academic and industrial.
■ Facilitate inter-disciplinary and inter-sectoral research collaborations, to develop the next generation of EV based research programmes.
■ Share best practice and provide a platform for collaborative rather than competitive bids, strengthening submissions to research councils for funding.
■ Train, support and inspire the next generation of outstanding leaders in EV research.
Then come along to the free MI Extracellular Vesicles Conference 2026!
This will be an in-person one day conference bringing together academics, local biotech and instrument manufacturers to discuss all things extracellular vesicles Come along to learn about the technology, hear something new in the field or present your research.
This conference is open to all researchers with an interest in extracellular vesicles, including those wanting to learn more about this exciting and rapidly expanding field. All are welcome, including Principal Investigators, Post-doctoral scientists, Technicians, Early Career researchers (including PhD / Masters students) and Industry representatives.
For sponsorship opportunities please email ellen.mcghee@midlandsinnovation.org.uk
Professor Lucy Fairclough – University of Nottingham and Chair of the Midlands Innovation Extracellular Vesicles Network
Artemis Stamboulis (University of Birmingham) – Exosome-Mediated Osteoblast–Osteoclast Crosstalk on Orthopaedic Implant Materials ABSTRACT 1
Anto Praveen Rajkumar Rajamani (University of Nottingham) – BBDLB: A novel multiplex plasma extracellular vesicle RNA biomarker assay for improving clinical diagnosis of Dementia with Lewy Bodies ABSTRACT 2
Maria Pia Savoca (Nottingham Trent University) – The zeta potential of urinary extracellular vesicles as a possible predictor of kidney disease in diabetes ABSTRACT 3
Adam Davison (Cytek) – Characterisation & Sorting (yes, you read right) of EVs using FSP technology and Imaging Flow’
Stephen Price (Beckman Coulter) – Nano sized wonders with CytoFLEX
Jessie Xu (Upper Biotech) – Turning EVs into Answers: Powering Biomarker Discovery with ExoFaster-500
Sophie McCann (University of Nottingham) – Investigating the role of microRNAs in medulloblastoma-to-neurone extracellular vesicle-mediated signalling ABSTRACT 4
Martina Gabrielli (Nottingham Trent University) – Microglial Extracellular Vesicles Promote Synaptic Pruning via C1q Deposition at the Synapse ABSTRACT 5
Charlie Mafham (University of Birmingham) – Epithelial derived extracellular vesicles – implications for immune mediated fibrotic lung disease ABSTRACT 6
Bev Goward (SONY) – Ensuring consistent results in Spectral Flow Cytometry across analysis and sorting platforms
Mel Disher (Analytik) – From Isolation to Insight: Integrating Exodus EV Isolation with ZetaView® Evolution Nanoparticle Tracking
Xiangru Huang (BGI Genomics) – Quality Multi-omics Service Provider Empower Your Research
Katherine White (University of Nottingham) – Impact of ALS-associated FUS dysfunction on extracellular vesicle-mediated miRNA signalling ABSTRACT 7
Junpeng Luo (University College Dublin) – Upstream Process Development Enables Scalable and Reproducible Manufacturing of Mesenchymal Stem Cell-derived Extracellular Vesicles ABSTRACT 8
Atiyeh Asadpour (University of Reading) – Neural Crest-Derived Stem Cell Secretomes Disrupt Glioblastoma through Dual-Pathway Inflammatory Rebalancing ABSTRACT 9
Yu-Su Chen (Malvern Panalytical) – Beyond size: exploring EVs through multimodal characterization
Tushar Shah (Secretech) – Proteomic Profiling of Single EVs Using Proximity Barcoding
P.j. Chana (Slingshot Biosciences) – Cell and EV mimics – A new era of controls
Professor Charlotte Lawson (University of Lancashire) – Functional effects of dietary derived extracellular particles on human inflammatory responses in vitro
Artemis Stamboulis, Sanjana Vig, Esra Cansever Mutlu, Maria Helena Fernandes*
Extracellular vesicles (EVs), particularly exosomes, play a crucial role in intercellular communication within the bone microenvironment and are increasingly recognised as regulators of osteoblast–osteoclast (OB–OC) crosstalk. Elucidating their role is essential for the development of orthopaedic implant materials with optimised osteogenic and antibacterial properties. This study investigated the functional role of EVs in osteogenic and osteoclastic differentiation and assessed material-dependent effects on EV characteristics and cargo.
Bone marrow–derived mesenchymal stem cells (BMSCs) and blood-derived monocytes were differentiated into osteoblasts and osteoclasts, respectively. EV biogenesis and secretion were modulated using GW4869, an inhibitor of ceramide-dependent exosome formation, and Nexinhib, an inhibitor of Rab27a-mediated vesicle exocytosis. EVs were isolated using ultracentrifugation and characterised by nanoparticle tracking analysis and transmission electron microscopy. The influence of metallic implant materials, including titanium, stainless steel, and TiAl6V4, on EV release was assessed during osteogenic differentiation under EV-depleted conditions.
GW4869 effectively reduced the release of small EVs from BMSCs in a dose-dependent manner and was associated with reduced alkaline phosphatase activity, indicating impaired osteogenic differentiation. In contrast, Nexinhib induced significant cytotoxicity in BMSCs, highlighting the importance of EV secretion for osteoblast viability. During osteoclast differentiation, both inhibitors altered EV size distribution and concentration, while unexpectedly enhancing tartrate-resistant acid phosphatase activity, suggesting a complex regulatory role of EVs in osteoclastogenesis. Material-dependent differences in EV size and concentration were observed, with a progressive reduction in EV output as osteogenic differentiation advanced.
Functional assays demonstrated that osteoclast-derived EVs significantly enhanced osteogenic differentiation of BMSCs in a dose- and stage-dependent manner, even in the absence of osteogenic supplements. Proteomic profiling of osteoclast-derived EVs revealed enrichment of cytoskeletal, metabolic, and signalling proteins associated with bone remodelling pathways. These findings demonstrate the central role of EV-mediated OB–OC communication and highlight the influence of implant materials on EV-mediated signalling.
Background:
Distinguishing Dementia with Lewy Bodies (DLB) from Alzheimer’s Disease (AD) accurately is essential because they differ in their prognosis, risk profiles, treatment, and sensitivity to antipsychotic medications. As DLB-specific imaging biomarkers and lumbar puncture are not feasible in many clinical settings in UK, there is an urgent clinical need for identifying blood-based biomarkers for DLB. We have reported DLB-specific RNA that were significantly differentially expressed in both post-mortem DLB brains and blood-based small extracellular vesicles (SEV) in people living with DLB. We aimed to develop a multiplex plasma SEV RNA biomarker assay using these DLB-specific RNA, and to establish its diagnostic accuracy.
Methods:
We designed customised BBDLB (Blood-based biomarker for DLB) miRNA and mRNA assays using the NanoString nCounter® technology. We recruited people with DLB, AD, and people without cognitive impairment (N=90;n=30/group) from four NHS sites. We separated SEV from platelet-poor plasma using size exclusion chromatography, and characterised SEV using cryo-transmission electron microscopy. BBDLB assays measured DLB-specific plasma SEV RNA levels. Using clinical diagnoses of consultant old age Psychiatrists with DLB expertise as reference standard, we estimated the diagnostic accuracy of our BBDLB assays.
Results:
We demonstrated the feasibility of measuring very low-input plasma SEV RNA levels from clinically collected blood samples. The BBDLB miRNA assay could correctly classify 78.4% of people with DLB from AD with 91.3% sensitivity, 57.1% specificity, 2.13 positive likelihood ratio (PLR) and 0.15 negative likelihood ratio (NLR). The BBDLB mRNA assay could correctly classify 75.0% of DLB from AD with 70.0% sensitivity, 80.0% specificity, 3.50 PLR and 0.38 NLR.
Conclusion:
The BBDLB assay is the first blood-based diagnostic biomarker assay for DLB that has the potential to be adapted for routine use in the NHS settings. Enriching plasma SEV for neuronal and microglial origin by immunoprecipitation can improve the signal-to-noise ratio and diagnostic accuracy.
The BBDLB assay is the first blood-based diagnostic biomarker assay for DLB that has the potential to be adapted for routine use in the NHS settings. Enriching plasma SEV for neuronal and microglial origin by immunoprecipitation can improve the signal-to-noise ratio and diagnostic accuracy.
Melanie Tepus1, Maria Pia Savoca1, Elisabetta A. M. Verderio1,2
1 Centre for Systems Health and integrated Metabolic Research (SHiMR), School of Science and Technology, Nottingham Trent University, Nottingham, United Kingdom
2Department of Biological, Geological, and Environmental Sciences, Alma Mater Studiorum, University of Bologna, Bologna, Italy
Introduction
Urinary extracellular vesicles (uEVs) have diagnostic potential as a source for biomarkers, however, their physical properties such as the zeta potential (ZP) are less characterised. ZP of nanoparticles may reflect the properties of the cells from which they originate and could serve as an indicator of underlying pathological changes. We hypothesized that uEVs’ ZP could vary in diabetic kidney disease (DKD) compared with diabetes with no kidney disease, with the possibility of early DKD detection and stratification.
Methods
Cell-free urines, sourced from Sheffield Kidney Institute and Patras University Hospital, (Greece) with informed consent and uniformed standards were grouped in 3 cohorts: DKD (n=36), further stratified based on eGFR loss in stable (<2 ml/min/year) and progressive (>5 ml/min/year); diabetes with no CKD (n=16); healthy (18). Small and large uEVs were obtained by differential centrifugation and differences in ZP, size distribution, and concentration analysed using nanoparticle tracking analysis and TEM. uEVs markers were further characterized by immunoblotting and SP-IRIS/Exoview.
Results
A significant difference in large uEVs ZP was observed between the diabetes and the healthy cohort, and the diabetes and DKD progressive cohort, with a less negative ZP in diabetes. No differences were reported in small uEVs. We hypothesised that alterations in cell surface heparan sulphate (HS) proteoglycans occurring in diabetes (loss of negatively charged glycocalyx) and DKD (HS deposition underlying tissue fibrosis) may be reflected in the EVs surface, resulting in changes in ZP, mirroring HS content. Indeed, uEVs from DKD displayed more HS than uEVs isolated from diabetic subjects when evaluated by SP-IRIS/Exoview, trapping EVs via HS-binding. Comparative uEV proteomics revealed an increased presence of sulfated glycosaminoglycans lumican in DKD EVs compared to diabetes, reported to increase in DKD in early fibrosis.
Conclusions
The zeta potential of large uEVs could be predictive of the evolution of diabetes into DKD.
McCann, Sophie, University of Nottingham (School of Life Sciences)*; Jackson, Hannah, University of Nottingham (Children’s Brain Tumour Research Centre, School of Medicine); Hathway, Gareth, University of Nottingham (School of Life Sciences); Coyle, Beth, University of Nottingham (Children’s Brain Tumour Research Centre, School of Medicine); Dajas-Bailador, Federico, University of Nottingham (School of Life Sciences). *corresponding author: mbysm9@nottingham.ac.uk
Paediatric brain tumour survivors are more likely to experience major psychiatric and neurodevelopmental disorders than their peers. To date, little research has been done on the neurobiological consequences of experiencing a brain tumour during early development.
We examined the ability of medulloblastoma tumour cells (the most common malignant paediatric brain tumour) to affect the development of cortical neurones via extracellular vesicle (EV)-mediated signalling. EVs were isolated from three medulloblastoma cell lines (DAOY, D425 – derived from a primary cerebellar tumour and D458 – derived from the patient-paired recurrent CSF metastasis) and added to E16.5 mouse primary cortical neurones for 24 hours. As indicators of cortical development and network maturation, axon length and total neuronal arborisation length were measured.
D458 EVs increased both axon and total neuronal arborisation length in the cortical neurones. However, neither D425 nor DAOY EVs had any effect on either output. To investigate the mediators of this effect, next-generation sequencing (Illumina, SY-415-1002) and unsupervised analysis (Qiagen) of the D458 EV microRNA (miRNA) cargo were performed, and three highly abundant miRNA candidates were selected. miRNA power inhibitors (100nM) were added to cortical neurones for 24 hours alongside D458 EVs. Inhibiting hsa-miRNA-X significantly attenuated the D458 EV-mediated increase in axon length and total neuronal arborisation length, indicating a potential mechanism for the observed EV-mediated effect. Potential mRNA targets of hsa-miRNA-X were identified in the literature and by using miRDB target prediction. Expression of these mRNA targets was validated in the mouse primary cortical neurones using RT-qPCR.
This study is helping to define how miRNA-containing medulloblastoma-derived EVs can reshape neuronal development. The outputs of this study are highly relevant for improving long-term neurological outcomes in survivors and for identifying novel biomarkers.
Martina Gabrielli1,2, Giulia D’Arrigo2, Giulia Cutugno2, Maria Teresa Golia2, Francesca Sironi3, Marta Lombardi2, Roberto Frigerio4, Marina Cretich4, Elisabetta Battocchio2, Sara Francesca Colombo2, Cristiana Barone5, Emanuele Azzoni5, Roberta Ghidoni6, Caterina Bendotti3, Rosa Chiara Paolicelli7, Claudia Verderio2
1 School of Science and Technology, Centre for Systems Health and Integrated Metabolic Research (SHiMR), Nottingham Trent University, Nottingham, United Kingdom
2 Institute of Neuroscience, National Research Council of Italy, Via Raoul Follereau 3, 20854, Vedano al Lambro, Italy.
3 Research Center for ALS, Istituto di Ricerche Farmacologiche Mario Negri IRCCS, Via Mario Negri, 2, 20156 Milano, Italy.
4 National Research Council of Italy, Institute of Chemical Science and Technologies (SCITEC-CNR), Via Mario Bianco 9, 20131 Milan, Italy.
5 School of Medicine and Surgery, University of Milano-Bicocca, Monza, Italy
6 Molecular Markers Laboratory, IRCCS Istituto Centro San Giovanni di Dio Fatebenefratelli, 25125 Brescia, Italy.
7 Department of Biomedical Sciences, Faculty of Biology and Medicine, University of Lausanne, Lausanne, Switzerland.
The complement factor C1q is released by microglia, localizes on weak synapses and tags them for microglia-mediated synaptic pruning, a fundamental process for proper circuit refinement across early postnatal life, reactivated in neurodegeneration. However, how C1q tags synapses at specific times remains elusive. We explored the possible involvement of extracellular vesicles (EVs) released by microglia in C1q delivery to synapses designated for removal.
We optimized a protocol to extract large and small EVs from mouse brain tissue. By Western Blot, TRPS, single molecules arrays (SimoA) and confocal imaging, we measured the amount, cell source and complement cargo of EVs extracted from WT postnatal pups and adult mice brains, and from C9orf72KO adult brains displaying enhanced microglia-mediated pruning. Then, we performed in vitro and ex vivo studies to inquire microglial EVs involvement in C1q deposition to the synapse and synaptic engulfment, by increasing or reducing microglial EVs production through EVs supplementation, C9orf72 silencing or pharmacological inhibition (GW4869).
We show that C9orf72KO microglia releases more EVs carrying C1q, providing a link between microglial EVs production and synaptic removal. Moreover, we report that production of C1q carrying microglial EVs peaks during the pruning period in the early postnatal hippocampus, confirming a positive correlation between EVs production and synaptic removal. In neuron-microglia co-cultures, microglial EVs make preferential contacts with synapses, deliver C1q to pre-synapses that externalize phosphatidylserine and promote synaptic removal. Interestingly, C9orf72KO microglia engulf more synaptic terminals and decrease synaptic density to a greater extent compared to WT microglia whereas inhibition of EVs release by GW4869 restores normal pre-synaptic density, providing mechanistic evidence linking EVs release to synaptic remodeling.
This study identifies microglial EVs as delivery vehicles for C1q to synapses targeted for removal and implicates abnormal EVs production from microglia in both neurodevelopmental and age-related disorders characterized by dysregulated synaptic pruning.
Extracellular vesicle (EV) mediated cellular crosstalk has been implicated in the progression of idiopathic pulmonary fibrosis (IPF), however no mechanisms have been elucidated. We investigated the surface phenotype and cargo of EVs derived from IPF patients, and their effect on immune cell function and metabolism. EVs were isolated from platelet-free plasma and broncho-alveolar lavage (BAL) of IPF patients and age-matched healthy controls; phenotype was assessed using Exoview & nanoflow cytometry. EV microRNA cargo was investigated with small RNA sequencing and quantified and validated with RT-qPCR. Monocyte-derived macrophages (MDMs) from healthy volunteers and alveolar macrophages (AMs) from lobectomy patients were treated with EVs for 24 hours prior to metabolic profiling and flow cytometric assessment of efferocytosis & phagocytosis. EPCAM+/CD9+ EVs are enriched in IPF patients compared to age-matched healthy controls (p=0.0273). MDM treatment with IPF patient plasma EVs decreased efferocytosis (fc=0.36, p=0.0078) and increased phagocytosis. Plasma EVs from healthy age-matched controls were enriched with miR-485-5p compared to IPF patients, suggesting a protective role for this novel microRNA. Macrophage dysfunction is associated with a pro-fibrotic phenotype, thus modulation of plasma EV cargo may offer a novel therapeutic strategy for IPF patients.
Katherine White1, Daniel Scott1, Federico Dajas-Bailador1, Sebastien Serres1, Rob Layfield1
1School of Life sciences, University of Nottingham
Amyotrophic lateral sclerosis (ALS) is a neurodegenerative disease characterised by the progressive loss of upper and lower motor neurones (MNs). Mutations in Fused in Sarcoma (FUS) are associated with aggressive, juvenile-onset ALS. FUS encodes a predominantly nuclear RNA-binding protein that regulates RNA metabolism, including RNA transport and miRNA processing. Pathogenic FUS mutations can cause cytoplasmic mislocalisation where it forms aggregates, leading to neuronal death. Recent work has identified FUS as a key regulator of extracellular vesicle (EV)-mediated export of selected miRNAs (García-Martín et al., 2022), but this has not been explored in human disease-relevant cell types such as astrocytes and neurones.
In ALS, reactive astrocytes can in some contexts promote neurotoxicity exacerbating MN death. Astrocytes and neurones communicate bidirectionally, with astrocyte-derived EVs being one route thought to deliver neuroprotective molecules including miRNAs to neurones. We hypothesise that FUS dysfunction contributes to ALS pathogenesis through aberrant EV-mediated cell-cell miRNA signalling.
Small RNA sequencing of control human iPSC-derived MNs and their purified EVs revealed an enrichment of miRNAs with known FUS-binding motifs in EVs relative to whole-cell miRNAs. This enrichment was lost in MNs carrying the pathogenic FUS P525L mutation. Similar EV enrichment of FUS-binding motif miRNAs was observed in primary astrocytes, supporting a role for FUS in the selective sorting and loading of EV-associated miRNAs in ALS-relevant cell types. Future work aims to extend these findings to iPSC-derived astrocytes to generate an EV–miRNA atlas defining “inclusion” versus “exclusion” miRNA motifs and identifying how FUS mutations alter neural EV cargo composition.
1*Junpeng Luo, 1,2,3 Jessica Whelan, 1Susan McDonnell.
* lead presenter. junpeng.luo@ucdconnect.ie, UCD School of Chemical and Bioprocess Engineering, Ireland
1 UCD School of Chemical and Bioprocess Engineering, Ireland
2 UCD Conway Institute of Biomolecular and Biomedical Research, Ireland
3 Science Foundation Ireland Research Centre for Pharmaceuticals (SSPC), Ireland
Extracellular Vesicles derived from human Mesenchymal Stem Cells (hMSC-EVs) possess promising efficacy in immunomodulation and tissue regeneration. Yet, their clinical translation is bottlenecked by limited process scalability and a lack of robust manufacturing frameworks. Furthermore, current hMSC-EVs production often relies on ill-defined supplements which complicates regulatory approval.
Here, we present the development of a scalable three-dimensional (3D) process for manufacturing hMSC-EVs and compare its performance with that of a monolayer culture process. We also present the development of a chemically-defined medium to enable regulatory-compliant manufacturing of hMSC-EVs. The work highlights the single vesicle characterization using highly sensitive Nano-scale Flow Cytometry (nFCM).
The 3D process achieved a peak viable cell density of 3.25 ± 0.14 ×105 cells mL-1, a 3.2-fold improvement over the initial process. MSCs preserved the phenotypes defined by the ISCT guideline throughout cultures with 93.4 ± 0.3 % viability at the end of EV production. The 3D process yielded 5.10 ± 0.14 × 109 particles mL-1 of conditioned medium and a specific productivity of 6.83 ± 0.35 ×103 particles cell-1 day-1, representing 5.2-fold and 5.8-fold increases over the monolayer process, respectively. Both 2D- and 3D-EVs were positive for canonical tetraspanins CD9, CD63 and CD81, and MSC markers CD29, CD44, CD73, and CD90. Interestingly, the phenotypes of MSC-EVs did not always mirror those of their parental cells. Nano-FCM also revealed that MSC markers preferentially associated with EVs of a size over 100 nm, whereas tetraspanins displayed heterogeneous expression on EVs of all size. Growth factor screening indicated that a combination of Insulin-Transferrin-Selenium, bFGF, TGF-β1, albumin, and ascorbic acid collectively supported consistent MSC expansion in the absence of ill-defined supplements.
In conclusion, upstream process optimization effectively boosts MSC-EV titre and productivity, bridging the gap between manufacturing capacity and clinical demand. Transition to chemically-defined production enables MSC-EV manufactured under the Quality by Design (QbD) framework.
Atiyeh Asadpour, PhD in Pharmacology, University of Reading
Glioblastoma is the most aggressive primary brain tumour with a poor clinical outcome, even after surgery, radiotherapy, and chemotherapy. Their proliferation is facilitated by a pro-inflammatory tumour microenvironment. Changing pro-inflammatory environment through the immunoregulatory secretome of adult neural crest-derived stem cells (NCSCs) could be a possible method to make the tumour more susceptible to standard therapies. NCSC secretomes and small extracellular vesicles (sEVs) can provide a clinically scalable, cell-free approach to reprogram tumour inflammatory networks. Methods: Secretomes and sEVs from fully characterised NCSCs (three donors) were applied to human glioma cell lines. Functional effects were assessed via cytokine arrays, soft agar colony formation, temozolomide (TMZ) sensitivity assays, and immunocytochemistry. Results: NCSC-derived secretomes and sEVs interacted with glioma cells and suppressed TNF α-induced proliferation, migration, sphere growth, and viability. Treatment inhibited nuclear p65 translocation, redirected cytokine output toward an anti-tumour profile, and sensitised tumour colonies to TMZ. Conclusions: NCSC secretomes and sEVs synergise with TMZ to overcome Glioblastoma(GBM) chemoresistance through simultaneous NF-κB suppression and immunomodulation. This Good Manufacturing Practice (GMP)-compatible strategy integrates tumour microenvironment reprogramming.
Bahareh Parsi. Supervisors: Andrew Devitt & Ivana Milic.
School of Biosciences, Aston University, Birmingham. B4 7ET.
Wound treatment, costing the UK £8.3 billion annually, highlights the need for innovative therapies to enhance tissue repair. Wound healing is a complex, multi-phase process that depends on the coordinated migration of multiple cell types. In the early stages, macrophages drive the inflammatory response before transitioning to an anti-inflammatory, pro-repair phenotype, while fibroblasts contribute by synthesising extracellular matrix components and restoring tissue architecture. Extracellular vesicles (EVs) play an important role in this process by delivering bioactive cargo containing immunomodulatory components, as well as cytokines and growth factors that regulate inflammation and support tissue regeneration.
The aim of this research was to generate and characterise secretomes from stably transfected HEK293F cell lines, and to evaluate their effects on a fibroblast cell line migration and wound closure using an in vitro scratch assay. Macrophage chemotaxis in response to these secretomes was assessed using a Transwell migration assay. To investigate whether EV-associated cargo drives the observed biological effects, selective pharmacological inhibitors were applied in secretome-induced macrophage migration assays.
The results demonstrated enhanced macrophage migration in response to multiple HEK293F-derived secretomes and support a role for EV-associated cargo in modulating immune cell behaviour and highlight the therapeutic potential of secretome-based strategies to promote inflammation resolution and tissue repair in chronic wounds.
Mayur Shetty, Andrew Devitt and Mariaelena Repici.
School of Biosciences, College of Health & Life Sciences, Aston University, Aston Triangle, Birmingham, B4 7ET, UK.
Astrocyte senescence is increasingly recognized as a contributor to neuroinflammation and alters intercellular communication in the aging brain. While it is known that astrocyte senescence alters their secretory phenotype, also known as senescence associated secretory phenotype -SASP, its impact on extracellular vesicle (EV) biogenesis and cargo remains poorly understood.
We investigated how astrocyte senescence alters the characteristics of EV, aiming to identify astrocyte-derived EV (ADEV)-based biomarkers associated with senescence.
Senescence was induced in H4 neuroglioma cells through two independent methods: pharmacological treatment with Palbociclib (a CDK4/6 inhibitor) and ionizing radiation (DNA damage response). Successful senescence induction was confirmed by abnormal nuclear morphology, loss of Lamin B1, and positive staining for senescence-associated β-galactosidase (SA-β-gal). EV secreted in the medium by young and senescent astrocyte cultures were isolated by size exclusion chromatography (Izon qEV 70nm) and subsequently characterized by nano flow cytometry (NanoFCM). Senescent ADEV did not show differences in size distribution or in CD9, CD63 and CD81 compared to their young counterparts. Interestingly, GLAST positive EV showed a two-fold increase in senescent astrocytes.
Our findings demonstrate that astrocyte senescence induces specific and measurable EV changes. These data highlight the potential of ADEV as non-invasive biomarkers for early detection and progression monitoring in neurodegeneration.
Adams, Abigail Elizabeth¹; Milic, Ivana¹; Rothnie, Alice¹; Devitt, Andrew¹*. ¹Aston Institute for Membrane Excellence School of Biosciences, Aston University. *corresponding author: a.devitt1@aston.ac.uk.
Introduction
Extracellular vesicles (EVs) play a central role in immune cell communication, transferring bioactive proteins, lipids, and nucleic acids that regulate inflammatory control, cellular migration, and the immune response. ATP-binding cassette (ABC) transporters are well-established regulators of lipid and eicosanoid transportation, which are key mediators in initiating and regulating inflammatory signals and immune cell behaviour within the immune system. However, whether ABC transporters present on the EV surface contribute functionally to EV-mediated communication within the immune system remains poorly defined. Here, the aim is to explore whether EV-associated ABC transporters actively facilitate lipid and eicosanoid-dependent signalling and the potential impact on immune cell recruitment.
Methods
Mass spectrometry datasets profiling EV surface proteins from multiple immune cell types were analysed using FunRich to identify potential ABC transporter candidates for further research. ABCC1 was selected for this study as it was consistently detected on the surface of EVs across immune cell–derived populations. EVs will be isolated from Jurkat cells using size-exclusion chromatography (SEC), and surface expression of ABCC1 will be validated using specific antibodies. Migration assays using THP-1 monocyte-derived macrophages will be used to evaluate the ability of isolated EVs against the secretome to induce chemotactic responses. Applying ABCC1 inhibitors will determine whether transporter activity is required for EV-driven immune cell migration and whether EV-associated transporter activity facilitates the transfer of lipid mediators between EVs and recipient cells.
Conclusion
This work will determine whether ABC transporters function as active components of the EV surface, regulating lipid and eicosanoid-dependent signalling within the immune system. By defining whether there is a link between EV surface transporter activity and inflammatory signalling, the study will advance our understanding of how EVs coordinate immune communication and identify specific ABC transporters as potential modulators of EV-driven inflammatory responses.
Abdallah Swaid1,2, Gabriel Landini1, Rasha Abu-Eid1
Extracellular vesicles (EVs) are key mediators of intercellular communication, facilitating the transfer of bioactive molecules between cells and their surrounding microenvironment. Increasing evidence indicates that EVs contribute to cancer progression and immune suppression, highlighting their potential as biomarkers and therapeutic targets in head and neck cancer (HNC). However, variability in EV isolation methods remains a major challenge, as it can significantly influence EV yield, purity, and biological functionality.
This study aimed to compare four EVs isolation techniques from HNC cell lines and normal oral keratinocyte cells. Ultracentrifugation (UC), ultrafiltration (UF), polymer-based precipitation (T), and a combined precipitation and ultrafiltration (T+UF) approach were assessed. Isolated EVs were characterised using nano-flow cytometry for size and concentration, transmission electron microscopy for morphology, bicinchoninic acid assay for protein quantification, and Fourier-transform infrared spectroscopy for biochemical profiling. Functional effects were assessed using wound scratch and cell proliferation assays.
UF produced the highest EV yield, followed by the combined T+UF method and UC, while precipitation alone resulted in the lowest yield. EV size distributions were comparable across all methods, although UF-isolated EVs were slightly larger on average. Notably, the T+UF approach provided the most favourable balance between EV yield and purity. Functional assays demonstrated that EVs isolated using precipitation alone exhibited reduced biological activity, whereas EVs isolated by UF, UC, and T+UF showed greater effects on cell migration and cell proliferation.
Overall, these findings demonstrate that EV isolation strategy significantly impacts both the physical characteristics and functional properties of EVs. The combined precipitation–ultrafiltration approach offers an efficient and reproducible method for isolating functional EVs with acceptable yield and purity.
Author
Abstract
Rebecca Pope, Gareth Hathway, Federico Dajas-Bailador
School of Life Sciences, University of Nottingham, Nottingham, NG7 2UH, United Kingdom.
Dorsal root ganglion (DRG) sensory neurones exhibit a distinctive pseudounipolar morphology, with a single axon that bifurcates to form peripheral and central terminals. These terminals occupy unique microenvironments and interact with diverse cell types and mediators. Such compartmentalised signalling is central to pain processing, yet the contribution of non-neuronal cells at the central terminal is incompletely understood. For example, how astrocytes and microglia influence DRG excitability is poorly defined.
To model the perisynaptic environment of central DRG terminals, we established mixed glial cultures derived from E16.5 murine spinal cord, comprising astrocytes and microglia. Exposure of these cultures to a pro-inflammatory cytokine cocktail (TNF-α, IL-1β, and C1q) generated extracellular vesicles (EVs) with sensitising capacity. When applied to DRG neurones for 24 hours, these EVs significantly enhanced neuronal excitability (p = 0.0036), evidenced by enhanced calcium responses to TRPV1 activation, measured in neuronal somata. This indicated a glia-driven EV-mediated sensitisation process.
To investigate the underlying molecular mechanism, we focused on the miRNA cargo of inflammatory glial EVs. Candidate miRNAs were compiled from published datasets of EVs derived from astrocytes and microglia under pro-inflammatory conditions. Three were selected for functional screening. Inhibition of a single candidate miRNA, achieved using a potent antisense inhibitor directed against the transcript, prevented EV-induced sensitisation in DRG neurones, thereby establishing a causal role for miRNA-mediated glia-neurone signalling.
These findings demonstrate that inflammatory glial EVs act as vehicles of intercellular communication that drive neuronal hyper-responsiveness at the central terminal. Moreover, they identify miRNAs as critical functional cargoes capable of shaping nociceptive signalling. Importantly, the ability of miRNA inhibitors to abolish EV-induced sensitisation highlights their translational potential as therapeutic tools to modulate pain-related plasticity. By defining a novel glia-to-neurone communication pathway, this work advances our understanding of central sensitisation mechanisms and suggests new avenues for the treatment of chronic pain.
Eleanor Stanton¹*, Beth Coyle¹, Jason Adhikaree¹
¹Biodiscovery Institute, University of Nottingham, UK
Triple-negative breast cancer (TNBC) is the most aggressive breast cancer subtype, characterised by early recurrence and lack of targeted therapies. Although chemo-immunotherapy has improved standard-of-care treatment, 15-30% of patients still relapse. A crucial next step in TNBC is to identify reliable biomarkers of therapy response and relapse. Extracellular vesicles (EVs) are increasingly recognised as mediators of treatment resistance and pre-metastatic niche formation through the transfer of oncogenic proteins and microRNAs, making them promising candidates as reliable biomarkers for treatment monitoring and metastatic risk. Prior investigation in our research group demonstrated differential EV miRNA expression between primary and recurrent cell lines, supporting a role for EVs as biomarkers of disease progression.
This project aims to isolate and characterise EVs and their RNA profiles in TNBC pre- and post- chemo-immunotherapy to identify potential candidates of therapy resistance and increased aggressiveness. Currently, feasibility has been established by successful EV isolation and validation from two TNBC cell lines, RNA extraction from pre-treatment EVs, and drug sensitivity assays to determine a clinically representative synergistic chemotherapy ratio to create a post- treatment model. Several TNBC patients have been recruited, blood samples have been obtained at diagnosis, and plasma extracted for subsequent EV- associated RNA extraction.
Moving forward, post-treatment resistant cell populations will be modelled in vitro, to begin comparison between conditions and investigate persister EVs. EV isolation, RNA extraction and Nanopore sequencing will be repeated to identify differentially expressed RNA candidates. In parallel, additional patient samples will be analysed at diagnosis, and following 3 months of chemo-immunotherapy treatment, to identify longitudinal EV-RNA alterations.
We hypothesise that EV-associated RNA profiles will differ following treatment exposure in cell lines and patient plasma samples, reflecting how EV cargo is remodelled with therapy pressures. Overall, aiming to further the understanding of EV-mediated resistance and metastatic mechanisms in TNBC, explore potential as a minimally invasive biomarker.
Yasmin Azizbayli1, Laura G. Bartlett2, Adam J. Watkins3, Amanda Tatler4, Victoria James5, and Lucy C. Fairclough1
1School of Life Sciences, University of Nottingham, Nottingham, UK.
2 Institute of Life Sciences, University of Southampton, Southampton UK.
3 School of Medicine and Population Health, University of Sheffield, Sheffield, UK.
4School of Medicine, University of Nottingham, Nottingham, UK.
5School of Veterinary Sciences, University of Nottingham, Nottingham, UK.
Email: yasmin.azizbayli@nottingham.ac.uk
Background: Extracellular vesicles (EVs) are key mediators of intercellular communication and play important roles in reproductive processes, including gametogenesis, fertilisation, early embryo development, and implantation. EVs also contribute to immune modulation within the reproductive environment. Immune cells, particularly macrophages, are critical regulators of gonadal development. Cigarette smoke adversely affects reproductive organs and is now a major contributor to infertility worldwide. However, the role of EVs in driving these mechanisms is unknown.
Methods: Time-mated pregnant C57BL/6J mice carrying embryonic day 16.5 (E16.5) pups were obtained from Charles River (UK). Embryonic gonads were dissected and cultured on agar blocks for 24–216 hours before enzymatic digestion in RPMI-1640 (phenol red–free) supplemented with 25 mM HEPES, 2% FBS, 1 mg/mL collagenase IV, and 100 µg/mL DNase I to generate single-cell suspensions. Cells were stained and analysed by spectral flow cytometry using a Sony ID7000 with markers for major immune cell populations. Gonads were also cultured in six ex-vivo media formulations to assess viability and EV staining. After culture, supernatants were centrifuged (300 g, 8 min), incubated with 10 µM calcein AM (Cat. no. 425201, BioLegend) for 1 hour at 37 °C to stain EVs, diluted in filtered PBS, and analysed on an ImageStreamX MkII (Amnis).
Results: Results showed that immune cells (CD45⁺) comprised approximately 3–7% of the fetal gonad. Among these, macrophages (F4/80⁺) were the predominant population, whereas T cells (CD3⁺) were the least abundant. For ex vivo culture, EV-depleted medium was optimal, maintaining approximately 98% cell viability after 216 hours of culture.
Conclusions: We have optimised protocols for flow cytometry panels and ImageStreamX analysis to examine immune cell and EV populations in reproductive organs. Future work will investigate the effects of CSE on immune cell and EV proportions in reproductive tissues, as well as assess gamete quality and foetal reproductive health.
Authors: Kieran, Sasha, Aston Institute for Membrane Excellence, School of Biosciences, Aston University; Wolffsohn, James, College of Health & Life Sciences Aston University; Devitt, Andrew, Aston Institute for Membrane Excellence School of Biosciences, Aston University*. *corresponding author: a.devitt1@aston.ac.uk.
Introduction
As we age, our ability to repair damage and reduce inflammation is impaired, increasing the likelihood of chronic wounds. Studies show that extracellular vesicles (EVs) can promote wound healing and resolve inflammation though the mechanisms remain ill-defined. We aim to define the role of EVs and their cargo on wound healing, focusing on ocular and dermal models.
Methods and results
A model of wound healing was formed and optimised using corneal epithelial and human dermal fibroblasts cell lines to produce a physiologically relevant 2D-cultured model using an automated approach, increasing output capabilities and reducing variability. To assess model validity, the effect of osmolarity changes (a characteristic of dry eye conditions) and UV exposure on wound repair was monitored. Complete closure occurred in corneal and dermal models by 48h and osmolarity changes and UV together significantly inhibited wound repair. Following validation, the impact of EV-containing secretome from HEK293F cells transfected with important immune controlling cargo was investigated to begin ascertaining significant molecules involved in EV-modulated wound healing.
Conclusion
In conclusion, a cellular wound healing model was optimised to compare corneal and dermal wound healing in healthy untreated cell lines and validated using external factors known to reduce the ability of repair. Following work will investigate the impact of EV from an increased variety of host cells including primary stem and immune cells. Further investigation is necessary to understand how EVs and their components may influence this mechanism, potentially leading to essential research in improving wound healing capabilities for all.
Shaun Hanley, Aston University; Andrew Devitt, Aston University; Ivana Milic, Aston University*. *corresponding author: i.milic@aston.ac.uk.
Background
Glioblastoma is the most common and aggressive primary brain cancer in adults and has poor prognosis and survival rates. Current treatment for glioblastoma consists of concurrent radiotherapy and chemotherapy with temozolomide (TMZ). A major feature in glioblastoma is the immunosuppressive tumour microenvironment, partly mediated by glioblastoma-derived extracellular vesicles (GBM-EVs). Proteins and miRNAs are often investigated when identifying immunomodulatory targets in GBM-EVs but there are many bioactive lipids which are immunomodulatory and are often not investigated so their roles in GBM-EV mediated immunomodulation are poorly understood. Therefore, we aimed to perform omics investigations into GBM-EV cargo and how it changes upon treatment with TMZ.
Methods & Results
A172 glioblastoma cell viability in response to TMZ treatment was assessed by MTT and flow cytometry. GBM-EVs were isolated from the conditioned media of TMZ-treated and control A172 cells by size exclusion chromatography. Physical characterisation of EVs was performed by nano-flow cytometry. Lipid extraction was performed using a methanol/MTBE extraction method, analysed by mass spectrometry, and identified using LipidSearch 5. Proteins underwent in solution digest and peptide enrichment by SPE before mass spectrometry analysis using the Orbitrap Ascend. Relative protein quantification was performed using Proteome Discoverer and Progenesis QI for Proteomics.
Conclusions
This analysis will uncover changes in GBM-EV cargo in response to TMZ treatment. Further analysis will reveal bioactive cargo in GBM-EVs which may contribute to immunomodulation. Future work will investigate changes in functional immune responses to incubation with GBM-EVs. This may enable the identification of novel therapeutic targets in glioblastoma.
Michael Joseph 1, Elisa Tonoli 1, Zita Balklava 2, Gareth Cave 1, Elisabetta AM Verderio 1
1 School of Science and Technology, Nottingham Trent University, Nottingham.
2 College of Health and Life Sciences, Aston University, Birmingham.
Mesenchymal stem cell-derived extracellular vesicles (MSC-EVs) are an emerging cell-free alternative for stem cell-based regenerative therapy. The regenerative properties of the MSC secretome are attributed to the bioactive molecules found in the cargo of MSC-EVs; however, the key molecules and mechanisms involved in MSC-EV-mediated neuroprotection remain poorly understood.
Our research group isolated and characterised MSC-EVs from four different umbilical cord-derived MSC (UC-MSC) lineages (tissue donated by Anthony Nolan). Through comparative proteomics, transcriptomics and cytokine profiling, the specific molecular composition of UC-MSC-EV cargo was identified, using primary dermal fibroblasts as a mesenchymal cell type control (lacking neuro-regenerative capability of MSC). It was revealed that nine proteins and thirteen miRNAs, including anti-inflammatory miRNA (miR146a-5p) were specifically increased in the EV preparations of at least three different UC-MSC cell lines in comparison to fibroblast EVs. In an in-vitro model of mouse hippocampal neurons (DIV 5) exposed to Aβ-peptide, overexpression of miR146a-5p by cell transfection led to increased tubulin-III-beta staining and axonal preservation, as quantified via immunofluorescence, suggesting neuronal cell protection. In Caenorhabditis elegans, ortholog-gene knockouts (cho-1, pat-3, and mir-71) of a selection of the specific human MSC-EV molecules (CTL2, ITB1, and miR146a-5p respectively) significantly impaired motor functions in vivo. We conclude that the newly identified MSC-specific EV proteins and miRNAs could be responsible for the neuroprotective effect of UC-MSCs.
Acknowledgements:
ARUK-East Midlands pump priming grant and Eranda Rothschild Foundation.
Feryal Dabbagh1,2, Stephane Gross2 & Andrew Devitt1,2
A healthy pregnancy depends on specialized Decidual Natural Killer (dNK) cells. Unlike regular NK cells that attack foreign invaders, dNK cells have a “regulatory” role, meaning they promote maternal-foetal tolerance and help the placenta grow. Recent studies showed that extracellular Vesicles (EVs) derived from dNK cells play important roles in the communication of immune cells and trophoblast cells. Because primary dNK cells are difficult to isolate and maintain, we used the NK-92 cell line as a scalable alternative. Our goal was to shift these naturally aggressive cells into a dNK-like state by reducing their supply of IL-2, a growth factor that usually drives their killing ability.
To test this, we cultured NK-92 cells with three different levels of IL-2 (100, 50, and 25 U/mL). We validated the transition by measuring how well the cells killed trophoblasts, whether they helped trophoblast cells proliferate in co-culture, and by tracking changes in their surface markers using flow cytometry.
The results showed that reducing IL-2 to 25 U/mL successfully triggered a functional shift. These cells stopped attacking trophoblasts and instead began supporting their growth, mimicking the behaviour of dNK cells in the uterus. On a molecular level, the activating “killing” receptor (NKp30) decreased while the inhibitory receptor (NKG2A) remained stable. Notably, these effects were even stronger under hypoxia. This suggests that the low-oxygen environment of the early uterus acts as a key signal to “switch” these cells into their regulatory model.
In conclusion, combining low IL-2 with hypoxia effectively skews NK-92 cells toward a dNK-like phenotype. This validated model provides a reliable platform for harvesting dNK-like exosomes, which are essential tools for studying how the mother’s immune system and the placenta communicate to ensure a successful pregnancy.
Baker, Natasha, School of Biosciences, Aston University; Devitt, Andrew, Aston Institute for Membrane Excellence, School of Biosciences, Aston University; Bottrill, Andrew, Proteomics RTP, School of Life Sciences, University of Warwick; Zampronio, Cleidiane, Proteomics RTP, School of Life Sciences, University of Warwick; Gross, Stephane, School of Biosciences, Aston University *. *corresponding author: s.r.gross@aston.ac.uk.
Trophoblast-derived extracellular vesicles (EV) mediate maternal-fetal communication and immunomodulation during placental development. However, the role of S100P in this process remains unknown. S100P has been implicated in migration and invasion as well as immunosuppression in the tumour microenvironment and so may regulate similar functions in the placenta. Therefore, our aim was to investigate the role of S100P in EV production under hypoxia (1% O2) or normoxia (20% O2), and the involvement of these EV in macrophage recruitment.
EV from HTR-8 trophoblast cell lines with exogenous S100P expression were first collected and quantified via flow cytometry. S100P expression was linked to a ≥2-fold increase in the hypoxic EV yield, with no significant difference in normoxic conditions. As S100P has been shown to interact with the actin-myosin cytoskeleton, this finding could indicate that S100P triggers membrane blebbing.
Then, to determine the function of EV, macrophage transwell migration assays were performed whereby the trophoblast EV and soluble fractions were isolated and investigated as attractants. The soluble fraction induced significant migration after 6-hours, while EV-induced migration was variable and occurred earlier when observed. No significant difference between S100P or oxygen tension was determined.
LC-MS analysis of the soluble fraction revealed an extracellular matrix (ECM) associated proteins, indicating that ECM-derived components may contribute to macrophage recruitment independently of EVs. These findings uncouple trophoblast EV from macrophage recruitment, suggesting EV have an alternative role in the placenta.
Mayhew, Megan, Nottingham Trent University; Megram, Oliver, Nottingham Trent University; Roshan, Simmie, Nottingham Trent University; Smith, Marshall J, JEOL; White, Samuel J, York St John University; Wilson, Philippe B, York St John University; Hunt, John A, Medical Technologies Innovation Facility; De Girolamo, Luigi, Nottingham Trent University; James, Victoria, University of Nottingham, Hunter, Elena, Medical Technologies Innovation Facility*. *Corresponding author: elena.hunter@ntu.ac.uk.
Early detection of cancer remains a critical challenge in clinical oncology, with significant implications for patient outcomes. Extracellular vesicles (EVs), which carry metabolites reflective of the metabolic state of their parent cells, represent a promising source of minimally invasive biomarkers. Metabolic profiling of EVs may offer novel insights into cancer biology and biomarker discovery.
A systematic review was undertaken to evaluate current literature on EV-derived metabolomic profiles in cancer to identify potential biomarkers. A comprehensive search of PubMed, Scopus, and Web of Science was conducted using a defined strategy. Included studies compared EVs derived from patients with cancer or cancerous cell lines with their respective healthy controls using a metabolomics approach. Data on cancer type, sample source, EV isolation and confirmation techniques, metabolite extraction, analytical platform, and metabolites identified were extracted. Twelve studies met the inclusion criteria and collectively reported 1,602 identified metabolites. Of these, 333 metabolites were reported to be differentially regulated in EVs derived from patients with cancer, or conditioned media from cancerous cell lines and their respective healthy controls. Interpretation of the literature was limited by heterogeneity in sample sources, EV characterisation, isolation techniques, and metabolomic platforms, complicating inter-study comparisons.
Despite methodological variation, distinct EV metabolic signatures were consistently reported, highlighting their potential for cancer biomarker discovery. However, current findings remain preliminary and require validation in larger and clinically diverse cohorts. Future EV metabolomic studies should adopt standardised guidelines for EV characterisation, metabolite identification, and results reporting to ensure reproducibility and facilitate clinical translation.
Chiara Suanno1,2, Iris Aloisi1, Elisa Tonoli2, Maria Pia Savoca2, Clare Coveney2, David J. Boocock2, Martina Levantino1, Giampaolo Zuccheri3, Stefano Del Duca1, Elisabetta AM Verderio2
1. University of Bologna, Department of Biological, Geological, and Environmental Sciences, Bologna (Italy)
2. Nottingham Trent University, Interdisciplinary Biomedical Research Centre, School of Science and Technology, Nottingham (UK)
3. University of Bologna, Department of Pharmacy and Biotechnology, Bologna (Italy)
Until 2007, plants were believed uncapable of secreting extracellular vesicles (EVs) due to the presence of a cell wall. In the last twenty years, it was demonstrated that plants can externalise different types of EVs, including exosomes, for intercellular communication or as a defence mechanism. Plant-derived EVs (PDEVs) have since arisen growing interest for their potential as carriers for drug delivery, but little is still known about their biogenesis, secretion mechanism, and role in the plant kingdom. While EVs isolation from living plants proves complicated, pollen represents a promising model to study in vivo PDEVs secretion. Pollen is the male gametophyte of seed plants (Spermatophytes), a haploid, microscopic organism that can be cultured in vitro in a liquid medium. We have previously demonstrated that pollen releases EVs exclusively during germination, while hydration either in a humid chamber or in particle-free PBS does not promote EVs secretion, implying that they are part of a physiological process. Moreover, we have analysed the protein content of EVs from Actinidia chinensis (L.) pollen by RP-HPLC-ESI-MS/MS and western blotting, assessing the presence of (I) EVs common markers like Clathrin, tetraspanins, ESCRT-related proteins, and Hsp; (II) Ole e 1, described as a pollen-derived EVs molecular marker; and (III) a plant-homolog of ALIX (BRO1-domain containing protein) which was enriched in the EV fraction according to the western blot. We are currently optimising the isolation method for pollen EVs, in order to achieve a better compromise between yield and purity, as part of the EU-funded project “EXIT PLAN” (MSCA Global Postdoctoral Fellowship). For this purpose, we have compared different approaches – filtration, differential centrifugation, sucrose gradient separation, precipitation –, evaluating EV yield by nanoparticle tracking analysis (NTA) and EV purity by atomic force microscopy (AFM) and immunoblotting.
Dimitri Aubert, Vesiculab Ltd.
Extracellular vesicle (EV) research is rapidly expanding across biomarker discovery, functional biology, and therapeutic development, yet workflows remain limited by complexity, variability, and scalability challenges. Vesiculab develops integrated solutions designed to streamline EV research workflows, enhance performance, and enable the generation of larger, higher-quality datasets.
Our approach addresses four critical stages of the EV workflow. First, EV isolation and purification are simplified using Vesi-SEC, a size exclusion chromatography (SEC) spin column platform that enables rapid, reproducible separation of EVs from contaminants while preserving vesicle integrity. Second, EV detection and labelling are improved with Vesi-Dye LMB, a family of lipophilic membrane-binding dyes optimised for consistent staining and compatibility with downstream analytical techniques. Third, robust data acquisition is supported by Vesi-Ref, a set of reference materials designed for instrument calibration and gating standardisation, improving reproducibility and comparability across experiments and platforms. Finally, EV handling and storage are stabilised using Vesi-Safe, a preservation buffer formulated to maintain EV integrity during processing and long-term storage.
Together, these solutions reduce hands-on time, minimise variability, and improve throughput, enabling researchers to scale their studies while maintaining data quality. By addressing key bottlenecks across the EV workflow, from isolation to analysis and preservation, Vesiculab contributes to more reliable and standardised EV research practices.
Vesiculab remains committed to continuous innovation to further enhance EV research outcomes and support the evolving needs of the field. In parallel, the company actively engages with the UK and East Midlands EV research community, fostering collaboration and contributing to the advancement of best practices and translational applications.
Author
Abstract
