Depending on their cell hosts, EVs contain functional cargos, including proteins, nucleic acids, and lipids. In biology, EVs play crucial roles in cell-to-cell communication, maintaining cellular homeostasis, regulating tissue development, and modulating immune functions. Characterizing EV-derived molecules in biological and disease states would lead to the discovery of precise biomarkers. EVs share the cytoplasm of pathogenic cells and are present in extracellular fluids. For example, one can follow this concept to develop a liquid biopsy for detecting oncogene amplification status through EV-derived mRNA upregulation, e.g., MYCN-amplified neuroblastoma, in adjunct to the routine clinical workup for the risk classification and subsequent therapeutic regimens.
Extracellular vesicles have emerged as important mediators of intercellular communication, and have gained interest for therapeutic potential as treatment itself and as biomarker of several pathologies. Research covers a wide range of aspects from basics to manipulation for therapeutic purposes and immunomodulatory to regenerative potential. Exosomes are extracellular vesicles released from cells upon fusion of an intermediate endocytic compartment, multivesicular body, within plasma membrane.
Novel technologies, including those based on nanotechnology, show promise for the further development and clinical use of extracellular vesicles as liquid biomarkers. Development of biomarkers is a long and complicated process, and there are still not many biomarkers based on extracellular vesicles in clinical use. However, the knowledge acquired during the last decade constitutes a solid basis for the future development of liquid biopsy tests for prostate cancer. These are urgently needed to bring prostate cancer treatment to the next level in precision medicine.
EVs are membrane-bound nanoparticles shed from most if not all cells of the body, and carry DNA, RNA, protein, and lipids that reflect the identity and molecular state of their cell-of-origin. EVs can cross the blood–brain barrier and their release is upregulated in neoplasia. In this review, we describe the current knowledge of EV biology, the role of EVs in glioma biology and the current experience and challenges in profiling glioma-EVs from the circulation.
The emerging role of exosomes as a natural nano-vehicle, which were once thought of as cell’s disposal system, offers a big potential in therapy. They are virtually in every cell type of the body ranging from 30-120 nanometers in size. For example, exosomes released by stem cells may deliver a cargo that inhibits or promotes tumor growth, aid in healing and regenerations, and more. Those that are produced by immune cells are known to have regenerative and protective effects in conditions such as cardiovascular disease and stroke. This only means that different cell types secrete exosomes with unique molecular delivery that performs, specifically in a targeted area. This unique property is a key consideration for manufacturing exosomes for therapeutic use.
Exosomes are considered as potential biomarkers that can reflect information from their parent cell-associated cancer microenvironment. Recently, aptasensors have been widely used for cancer and tumor exosome detection. Aptamers related to exosome surface proteins are usually used to introduce a sequence; the aptamer is used for exosome recognition, and the introduced sequence is used to form G-quadruplexes and for signal amplification.
Cancer-cell-derived exosomes are regarded as noninvasive biomarkers for early cancer diagnosis because of their critical roles in intercellular communication and molecular exchange. A robust aptamer-initiated catalytic hairpin assembly (AICHA) fluorescence assay is proposed for universal, sensitive detection of cancer-derived exosomes.
As a subspecies of extracellular vesicles (EVs), exosomes have provided promising results in diagnostic and theranostic applications in recent years. The nanometer-sized exosomes can be extracted by liquid biopsy from almost all body fluids, making them especially suitable for mainly non-invasive point-of-care (POC) applications. To achieve this, exosomes must first be separated from the respective biofluid. Impurities with similar properties, heterogeneity of exosome characteristics, and time-related biofouling complicate the separation. This practical review presents the state-of-the-art methods available for the separation of exosomes. Furthermore, it is shown how new separation methods can be developed. A particular focus lies on the fabrication and design of microfluidic devices using highly selective affinity separation.
The small size and physicochemical heterogeneity of exosomes make their physical characterization and quantification an extremely challenging task. Creative Biolabs has successfully launched an innovative exosome quantification platform with multiple high-end technologies to detect and quantify exosomes in complex biological samples.
Biotium, a leading supplier and inventor of fluorescent cellular probes for life science research, announces the release of new ExoBrite™ Antibody Conjugates. These antibodies were curated specifically for optimal detection of exosome markers CD9, CD63, and CD81 in isolated exosomes or exosome extracts with little to no background. ExoBrite™ Antibody Conjugates are available in formulations for both flow cytometry and western blotting.
The exosome is considered a useful biomarker for the early diagnosis of cancer. However, pretreatment of samples used in diagnosis is time-consuming. Researchers at Kwangwoon University have fabricated a capacitance-based electrical biosensor that requires no pretreatment of the sample; it is composed of a DNA aptamer/molybdenum disulfide (MoS2) heterolayer on an interdigitated micro-gap electrode (IDMGE)/printed circuit board (PCB) system for detecting exosomes in an undiluted serum sample. The DNA aptamer detects the CD63 protein on the exosome as the biomarker, while the MoS2 nanoparticle enhances electrical sensitivity.
Size exclusion chromatography (SEC) has been described as most efficient method for isolating EVs from complex biological fluids in one single step, with a good recovery and with almost complete removal of contaminants, such as proteins and lipoproteins. Immunostep have developed 70nm and 35nm SEC columns for EVs isolation from complex biological fluids such as: plasma, serum, urine and cerebrospinal fluid.
Researchers from Duke University have developed an electrochemical micro-aptasensor for the highly sensitive detection of exosomes by integrating a micropatterned electrochemical aptasensor and a hybridization chain reaction (HCR) signal amplification method. Specifically, exosomes are enriched on CD63 aptamer-functionalized electrodes and then recognized by HCR products with avidin-horseradish peroxidase (HRP) attached using EpCAM aptamers as bridges. Subsequently, the current signal that is generated through the enzyme reaction between the HRP enzyme and 3,3’,5,5’-tetramethylbenzidine (TMB)/H2O2 directly correlates to the amount of bound HRP on the HCR products and thus to the number of target exosomes.
Proteomic profiling of extracellular vesicles (EVs) represents a promising approach for early detection and therapeutic monitoring of diseases such as cancer. The focus of this study was to apply robust EV isolation and subsequent data-independent acquisition mass spectrometry (DIA-MS) for urinary EV proteomics of prostate cancer and prostate inflammation patients. Urinary EVs were isolated by functionalized magnetic beads through chemical affinity on an automatic station, and EV proteins were analyzed by integrating three library-base analyses (Direct-DIA, GPF-DIA, and Fractionated DDA-base DIA) to improve the coverage and quantitation.
Extracellular vesicles (EVs) have attracted interest due to their ability to provide diagnostic information from liquid biopsies. Cells constantly release vesicles divers in size, content and features depending on the biogenesis, origin and function. This heterogeneity adds a layer of complexity when attempting to isolate and characterize EVs resulting in various protocols. Their high abundance in all bodily fluids and their stable source of origin dependent biomarkers make EVs a powerful tool in biomarker discovery and diagnostics. However, applications are limited by the quality of samples definition.
Mechanistically, we will analyze the implication of EVs in pathological processes associated to cardiovascular remodeling, with special emphasis in their role in vascular and valvular calcification. Specifically, we will focus on the participation of EVs in calcium accumulation in the pathological vascular wall and aortic valves, involving the phenotypic change of vascular smooth muscle cells (SMCs) or valvular interstitial cells (IC) to osteoblast-like cells. The knowledge of the implication of EVs in the pathogenic mechanisms of cardiovascular remodeling is still to be completely deciphered but there are promising results supporting their potential translational application to the diagnosis and therapy of different CVDs.
Extracellular vesicles (EVs) have shown great potential for biomarker profiling and disease treatment. The recent decade has witnessed rapid advancements in extracellular vesicle-based diagnosis and therapy. With the development of interdisciplinary approaches, biomedical engineering (BME) technologies play increasingly important roles in EVs research to facilitate the translation of EV study from bench to bedside, which may be realized by engineering EVs through multidisciplinary collaborations including medicine, biology, physics, chemistry, materials science and engineering, etc.
EXOSOMES are biological nano-vesicles involved in cell communication. Their discovery has revolutionized medicine. France is third in the world in terms of the number of clinical studies related to innovative diagnostics around these exosomes. Half of them take place in New Aquitaine. However, exosomes are still largely unexploited due to a lack of suitable tools allowing them to be characterized in real time according to their content. The ExoFlow project, funded by Nouvelle-Aquitaine region, aims to design and manufacture an innovative scientific instrument to characterize biological nano-objects such as exosomes or viruses in real time. The partners, ALPhANOV, Chrysa-link, the SATT Aquitaine deep tech incubator, the LP2N laboratory and the Paul Pascal Research Center will have 2 years to achieve these objectives.
Exosomes are first found in the supernatant of sheep red blood cells cultured in vitro. They are vesicles secreted by cells with relatively uniform size, a diameter of 30~160 nanometers and a density of 1.10~1.18 g/mL. Exosome carries a variety of proteins, mRNAs and miRNAs, and participate in the processes of cell communication, cell migration, angiogenesis and tumor cell growth, and may become a natural carrier of drugs for clinical treatment. Isolation and characterization of exosomes are technically challenging due to the persistence of contaminants such as microcapsules, lipoprotein particles, microorganisms, microbodies, protein aggregates, and DNA and other necrosis products during purification. Therefore, exosome characterization techniques are of critical importance in the exosome field.
The exosome is considered a useful biomarker for the early diagnosis of cancer. However, pretreatment of samples used in diagnosis is time-consuming. Researchers at Kwangwoon University have fabricated a capacitance-based electrical biosensor that requires no pretreatment of the sample; it is composed of a DNA aptamer/molybdenum disulfide (MoS2) heterolayer on an interdigitated micro-gap electrode (IDMGE)/printed circuit board (PCB) system for detecting exosomes in an undiluted serum sample. The DNA aptamer detects the CD63 protein on the exosome as the biomarker, while the MoS2 nanoparticle enhances electrical sensitivity.
“The Nanosight™ platform is an advanced unique platform that is established for exosome characterization. It’s made accessible to all kinds of research and industrial customers for the size distribution analysis, exosome concentration measurement, as well as the phenotyping evaluation.”
Precise detection of exosome concentration is difficult because it requires clinical sample pretreatment. To solve this problem, the use of the nanobiohybrid material-based biosensor provides improved sensitivity and selectivity. The present review will discuss recent progress in exosome biosensors consisting of nanomaterials and biomaterial hybrids for electrochemical, electrical, and optical-based biosensors.
Exosomes are a class of membrane secreted vesicles found in most bodily fluids. Typically 50–100 nm in size, they are involved in intracellular communication, metastasis, and antigen presentation, and carry important cellular information. They have become a source of increasing interest for potential novel biomarkers. However, exosomes can be difficult to characterize and study due to their small size. The ZE5 Cell Analyzer is ideally suited for studying them though as it can be equipped with a small particle detector (SPD) that uses the 405 nm laser line and a separate forward scatter (FSC) PMT to help detect exosomes and other sub-micron particles. It also has a novel threshold plot that provides a raw signal from the trigger detector(s) and aids in discriminating noise and debris from the desired signal.
Research on exosomes as a potential source of biomarkers of human disease has increased rapidly in recent years and consequently there are a large number of techniques for the isolation and characterization of this type of extracellular vesicles. However, many of the current techniques are poorly standardized. Furthermore, the use of exosomes in diagnostic tests or clinical research requires a sensitive, reproducible, and high-throughput method for the detection, characterization, and quantification of exosome samples.
Biotium, a leading supplier and inventor of fluorescent cellular probes for life science research, announces the release of new ExoBrite™ EV Membrane Staining Kits. These kits are designed for fluorescent staining of exosomes or extracellular vesicles (EVs) with little to no background. ExoBrite™ EV Membrane Stains are validated for flow cytometry, but also have potential applications in super-resolution imaging or other exosome detection platforms.
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