Project description:Crude extracellular vesicles (EVs) from eight healthy volunteers were separated into 6 fractions based on their densities by using the iodixanol-based density gradient centrifugation method. To determine the distribution of miRNAs among these fractions, quantities of 93 miRNAs were quantified by the TaqMan real time PCR method using the BioMark HD system (Fluidigm) equipped with 96.96 dynamic array (Fluidigm).
Project description:Crude extracellular vesicles (EVs) from eight healthy volunteers were separated into 6 fractions based on their densities by using the iodixanol-based density gradient centrifugation method. To determine the distribution of miRNAs among these fractions, quantities of 93 miRNAs were quantified by the TaqMan real time PCR method using the BioMark HD system (Fluidigm) equipped with 96.96 dynamic array (Fluidigm). Six samples were fractionated from a crude EVs by density gradient centrifugation. Total of 48 samples were prepared from 8 healthy volunteers. Technical replicate of 4 gave 8 x 6 x 4 x 93 = 17,856 data. As control Tris-HCl EDTA buffer (TE) was used.
Project description:Recent literature has documented the use of microRNAs (miRNAs) from circulating extracellular vesicles (EVs) as biomarkers for a plethora of diseases. The aim of this prospective study was to identify the diagnostic value of plasma EV-miRNAs in sepsis.Sepsis patients and healthy controls were matched for age and gender. EVs were separated from plasma of sepsis patients at admission as well as healthy controls. The expression of EV-miRNAs was evaluated by microarray and qRT-PCR.
Project description:Membrane vesicles released by neoplastic cells into extracellular medium contain potential of carrying arrays of oncogenic molecules including proteins and microRNAs (miRNA). Extracellular (exosome-like) vesicles play a major role in cell-to-cell communication. Thus, the characterization of miRNAs of exosome-like vesicles is imperative in clarifying intercellular signaling as well as identifying disease markers. microarray analysis identified several oncogenic miRNA between the two types vesicles.
Project description:Salivary extracellular vesicles (EVs) represent a powerful, non-invasive source of biomarkers for disease diagnosis and monitoring. Their molecular cargo reflects systemic and local physiological states, offering a window into neurological and inflammatory disorders. However, the diversity of EV isolation protocols and the possibility that each enriches distinct EV subpopulations remains a major barrier to reproducibility and data comparability. We conducted a comprehensive comparison of three EV isolation methods: ultracentrifugation (UC), PEG-based precipitation (Q), and immunoaffinity capture (M) to evaluate their impact on EV yield, purity, and molecular composition. Salivary EVs from healthy volunteers were analysed using proteomic and small-RNA sequencing approaches. Principal component analysis revealed clear isolation method-dependent clustering, where M-derived EVs displayed the most distinct profile. UC and Q produced broader proteomic repertoires with higher total protein content, whereas M-isolated EVs exhibited greater purity and enrichment of trafficking- and lysosome-associated proteins. Over 731 miRNAs selected, 28 were consistently altered across methods and 65 uniquely enriched in M isolates. RT-qPCR confirmed key directional trends. These 93 method-dependent miRNAs have predicted targets associated with synaptic structure and neurodegenerative pathways. These findings show that isolation methodology deeply shapes salivary EV cargo and suggest that immunoaffinity capture can isolate specific EV populations meeting diagnostic requirements.
Project description:MSCs from salivary gland or wharton's jelly were subjected to gene expression analysis, and their extracellular vesicles were used for exosomal miRNA profiling.
Project description:miRNA-sequencing of grapefruit-derived extracellular vesicles and fusion nanovesicles derived from grapefruit-derived extracellular vesicles and gingival mesenchymal stem cell-derived vesicles. We then performed gene expression profiling analysis to explore the miRNAs derived from grapefruit-derived extracellular vesicles, and the retention rate of miRNAs after membrane fusion
Project description:This dataset contains the single-cell gene expression profiles of mouse haematopoietic stem cell and progenitor cell populations isolated by FACS and paired gene expression profiles of their progeny (paired daughter cell assay and paired grand daughter cell assay). Expression of 96 TaqMan probes (A-MTAB-650) was assayed in individual cells by qRT-PCR using 96·96 Dynamic Arrays and the Biomark HD platform (Fluidigm).
Project description:Chronic pelvic pain syndrome (CPPS) and chronic prostatitis (CP) is difficult to distinguish from each other, herein termed CP/CPPS. This study aimed at gaining further insight into the change of extracellular vesicles (EVs) in prostatic fluid of male CPPS. From December 2019 to November 2020, after clinical screening, 24 patients with CPPS without obvious urinary symptoms and 13 healthy male participants were included. EVs were isolated from expressed prostatic secretion (EPS) of all subjects. The small non-coding ribonucleic acid (sncRNA) expression of EVs was sequenced, analysed, and validated by quantitative real-time polymerase chain reaction (qRT-PCR) assays. The results showed that plenty of sncRNAs were differentially expressed between the patients and healthy participants. Further qRT–PCR assays validated that several chronic pain-related miRNAs, including miR-204-5p, let-7d-3p, let-7b-3p, let-7c-3p, miR-146a-5p, and miR-320a-5p, were differentially expressed. Series sncRNAs including several chronic pain-related miRNAs were altered in EVs in prostatic fluid of patients with CPPS, which may serve as diagnostic markers for CPPS.