A5035909058- Extracellular vesicles in disease
- Genomic variations and chromosomal abnormalities
- Immunotherapy and Immune Responses
- T-cell and B-cell Immunology
- Chromosomal and Genetic Variations
- Lattice Boltzmann Simulation Studies
- Single-cell and spatial transcriptomics
- Immune Cell Function and Interaction
- Nanoplatforms for cancer theranostics
- Chronic Lymphocytic Leukemia Research
- Chronic Myeloid Leukemia Treatments
- MicroRNA in disease regulation
- Nanopore and Nanochannel Transport Studies
- RNA Interference and Gene Delivery
- Advanced biosensing and bioanalysis techniques
- Genetics and Neurodevelopmental Disorders
- Immune Response and Inflammation
- Cell Adhesion Molecules Research
- Microfluidic and Bio-sensing Technologies
- Musculoskeletal synovial abnormalities and treatments
- Acute Myeloid Leukemia Research
- Acute Lymphoblastic Leukemia research
- Nanoparticle-Based Drug Delivery
- Hematopoietic Stem Cell Transplantation
- Cancer Genomics and Diagnostics
Utrecht University
2014-2024
Wilhelmina Children's Hospital
2005
University of California, San Diego
2005
University Medical Center Utrecht
2005
Erasmus University Rotterdam
1995-1999
Netherlands Organisation for Applied Scientific Research
1990
Rotterdam University of Applied Sciences
1988
Leiden University Medical Center
1986
ABSTRACT Extracellular vesicles (EVs) are small, heterogeneous and difficult to measure. Flow cytometry (FC) is a key technology for the measurement of individual particles, but its application analysis EVs other submicron particles has presented many challenges produced number controversial results, in part due limitations instrument detection, lack robust methods ambiguities how data should be interpreted. These complications exacerbated by field's reporting framework, EV‐FC manuscripts...
Abstract Submicron‐sized vesicles released by cells are increasingly recognized for their role in intercellular communication and as biomarkers of disease. Methods high‐throughput, multi‐parameter analysis such extracellular (EVs) crucial to further investigate diversity function. We recently developed a high‐resolution flow cytometry‐based method (using modified BD Influx) quantitative qualitative EVs. The fact that the majority EVs is <200 nm size requires special attention with...
Essentials Platelet extracellular vesicles (EVs) concentrations measured by flow cytometers are incomparable. A model is applied to convert ambiguous scatter units EV diameter in nanometer. Most included lack the sensitivity detect EVs of 600 nm and smaller. The outperforms polystyrene beads for comparability platelet concentrations.Background Detection cytometry has poor interlaboratory comparability, owing differences cytometer (FCM) sensitivity. Previous workshops distributed set a...
Extracellular vesicles (EVs) in blood plasma are recognized as potential biomarkers for disease. Although is easily obtainable, analysis of EVs at the single particle level still challenging due to biological complexity this body fluid. Besides EVs, contains different types lipoproteins particles (LPPs), that outnumber by orders magnitude and which partially overlap biophysical properties such size, density molecular makeup. Consequently, during EV isolation LPPs often co-isolated....
Balanced chromosome rearrangements (BCRs) can cause genetic diseases by disrupting or inactivating specific genes, and the characterization of breakpoints in disease-associated BCRs has been instrumental molecular elucidation a wide variety disorders. However, mapping using traditional methods, such as situ hybridization with fluorescent dye-labeled bacterial artificial clones (BAC-FISH), is rather laborious time-consuming. In addition, resolution BAC-FISH often insufficient to unequivocally...
Many cell types release nanosized vesicles derived from endosomal compartments (exosomes) or the plasma membrane. Vesicles actively released by CD4(+) T cells have immune-modulatory characteristics. Using our recently developed high-resolution flow cytometry-based method for analysis of individual vesicles, we here investigated how receptor (TCR)-triggering and co-stimulatory signals influence quantity characteristics cells. We found that number within buoyant density range characteristic...
Essentials Extracellular vesicles (EVs) in biological fluids are promising biomarkers for disease. Fluorescence-based flow cytometric analysis is suitable to detect low abundant EV subsets. Particles of non-interest can induce false-positive light scatter and fluorescent signals. Interference particles be monitored by analyzing serial dilutions.Background plasma increasingly being recognized as potential biomarkers. diagnostic purposes should robust allow subsets with a wide range abundance...
Several naked virus species, including members of the Picornaviridae family, have recently been described to escape their host cells and spread infection via enclosure in extracellular vesicles (EV). EV are 50–300 nm sized lipid membrane-enclosed particles produced by all that broadly recognized for playing regulatory roles numerous (patho)physiological processes, viral infection. Both pro- antiviral functions ascribed released virus-infected cells. It is currently not known whether this...
Abstract Mast cells (MC) are well known for their effector role in allergic disorders; moreover, they associated with diverse modulatory effects innate and adaptive immunity. It is largely unclear how MC exert these modulating functions. In this article, we show that IgE-mediated degranulation leads to a rapid release of high quantities extracellular vesicles (EV), comparable the preformed mediators. EV submicron structures composed lipid bilayers, proteins, nucleic acids released by...
Nano-sized membrane vesicles are secreted by many cell types. These can serve as carriers of cellular information. DC-derived be targeted to other immune cells and modify their function. Accurate analysis quantitative qualitative changes in EV production DC upon different activation stimuli is needed further reveal the regulatory properties EVs. However, methods for reliable quantification individual EVs heterogeneity populations limited. With our recently developed high-resolution flow...
Previous studies have suggested that murine peritoneal cavity-derived B-1a cells possess similarities with described regulatory B cell subsets. The aim of the current study was to examine potential immunoregulatory function B(-1a) cells. In vitro activation B- and shows these anti-CD40 LPS induces secrete more IL-10, IL-6 IgM as compared splenic a suppression assay, CD40/TLR4-activated cavity functions they inhibit capacity CD4+ T produce both tumor necrosis factor-α interferon-γ. Splenic...
Abstract Capacitation is the final maturation step spermatozoa undergo prior to fertilisation. The efflux of cholesterol from sperm membrane extracellular environment a crucial during capacitation but current methods quantify this process are suboptimal. In study, we validate use BODIPY-cholesterol assay in vitro capacitation, using boar as model species. novel flow cytometric was validated with endogenous loss measured by mass spectrometry and compared filipin labelling. Following exposure...
Research in chickens has been fundamental for the discovery of basic aspects immune system and led to an interest in-depth characterization avian cell types including dendritic cells (DCs). The vitro generation expansion chicken bone marrow-derived (chBMDCs) presence GM-CSF provided a way study DCs, which are only present at limited numbers vivo. This method employed interactions between DCs pathogens or vaccines. However, detailed chBMDC culture was still lacking. In study, we performed...
A muscle progenitor cell population, other than satellite cells, can be isolated and purified from porcine tissue. We show the presence of at least two types stem cells in muscle: those that express α6 integrin lack expression this type. By flow cytometry, we could select for myogenic populations expressing neural adhesion molecule absence integrin. The showed an advantage formation myotubes, possibly by improved fusion capacity. This notion was strengthened qRT-PCR analysis showing...
Extracellular vesicles (EVs) in synovial fluid (SF) are gaining increased recognition as important factors joint homeostasis, regeneration, and biomarkers of disease. A limited number studies have investigated EVs SF samples patients with disease, but knowledge on the role healthy joints is lacking. In addition, no standardized protocol available for isolation from SF. Based high viscosity caused by concentrations hyaluronic acid (HA) – a prominent extracellular matrix component it was...
The long-term prognosis after surgical resection of malignant insulinoma (INS) is poor. Novel adjuvant therapies, specifically targeting cancer stem cells (CSCs), are warranted. Therefore, the goal this study was to characterize and target putative INS CSCs. Using fluorescence-activated cell sorting, human line CM pancreatic carcinoid BON1 were screened for presence cell-associated markers. CD90, CD166, GD2 identified as potential CSC Only CD90(+) had an increased tumor-initiating in athymic...
Abstract Flow cytometry allows multiparameter analysis on a single‐cell basis and is currently the method of choice to rapidly assess heterogeneity cell populations in suspension. With research field extracellular vesicles (EV) expanding, there an increased demand address EV biological samples. Although flow would be ideal technique do so, available instruments are general not equipped optimally detect dim light scatter signals generated by submicron‐sized particles like EV. sideward...
Abstract Fluorescent in situ hybridization allows for rapid and precise detection of specific nucleic acid sequences interphase metaphase cells. We applied fluorescent to human lymphocyte nuclei suspension determine differences amounts chromosome target amongst individuals by dual beam flow cytometry. Biotinylated 1 Y repetitive satellite DNA probes were used measure polymorphism eight healthy volunteers. The probe fluorescence was found vary considerably male volunteers (mean 169, S.D....