Isolation and characterization of extracellular vesicles

2019 ◽  
Vol 1 (1) ◽  
pp. 18-26
Author(s):  
Fabia Fricke ◽  
Dominik Buschmann ◽  
Michael W. Pfaffl
Keyword(s):  
Cell Culture ◽  
Extracellular Vesicles ◽  
Culture Media ◽  
Body Fluids ◽  
Cell Culture Media ◽  
Advantages And Disadvantages ◽  
The Past ◽  
Isolation And Characterization ◽  
Basic Biology

Research into extracellular vesicles (EVs) gained significant traction in the past decade and EVs have been investigated in a wide variety of studies ranging from basic biology to diagnostic and therapeutic applications. Since EVs are secreted by most, if not all, eukaryotic and prokaryotic cells, they have been detected in body fluids as diverse as blood, urine and saliva as well as in cell culture media. In this chapter, we will provide an overview of EV isolation and characterization strategies and highlight their advantages and disadvantages.

Characterization of the impact of classical cell‐culture media on the response of electrochemical sensors

2021 ◽  
Author(s):  
Ayman Chmayssem ◽  
Lauriane Petit ◽  
Nicolas Verplanck ◽  
Véronique Mourier ◽  
Séverine Vignoud ◽  
...  
Keyword(s):  
Cell Culture ◽  
Electrochemical Sensors ◽  
Culture Media ◽  
Cell Culture Media ◽  
The Impact

scholarly journals Isolation of Extracellular Vesicles from Cell Culture Media by Differential Ultracentrifugation  v1 (protocols.io.bnr3md8n)

protocols.io ◽  
2020 ◽  
Author(s):  
Dima Ter ◽  
Wendy Trieu ◽  
Maia Norman ◽  
Roey Lazarovits ◽  
George Church ◽  
...  
Keyword(s):  
Cell Culture ◽  
Extracellular Vesicles ◽  
Culture Media ◽  
Cell Culture Media

Characterization of Biomimetic Mineral Coated 3D PLGA Scaffolds

2006 ◽  
Author(s):  
A. Champa Jayasuriya ◽  
Chiragkumar Shah ◽  
Vijay Goel ◽  
Nabil A. Ebraheim
Keyword(s):  
Cell Culture ◽  
Culture Media ◽  
Biological Molecules ◽  
Polymer Surfaces ◽  
Human Blood Plasma ◽  
Carbonate Apatite ◽  
X Ray Diffraction ◽  
Cell Culture Media ◽  
Salt Leaching

The bone-like carbonate apatite (BLCA) coatings can be coated biomimetically in the polymer surfaces by soaking in the simulated body fluid (SBF). This SBF contains similar ionic constituents to human blood plasma. Micro-porous 3D poly(lactic-co-glycolic acid) PLGA scaffolds were fabricated by the solvent casting/salt leaching technique using chloroform to dissolve the polymer. We accelerated the deposition of mineral on scaffolds for 1-2 days, modifying the mineralization process using surface treatments and 5x SBF. These scaffolds were analyzed by Scanning Electron Microscopy (SEM), Fourier Transform Infra-Red (FTIR) and X-ray Diffraction (XRD). The scaffolds coated with BLCA layer were placed in the 24 well plates containing 2 ml of media, such as Tris Buffered Saline-pH 7.4, cell culture media containing αMEM supplemented with 10% FBS, and 1% penicillin-streptomycin and incubated at 37°C for 21 days. The BLCA layer on surfaces of scaffold was stable even after 21 days immersed in Tris Buffered Saline and cell culture media. This study suggests that BLCA were stable for at least 3 weeks in the both media, and therefore, mineral has a potential to use as a carrier for biological molecules for localized release applications as well as bone tissue engineering applications.

scholarly journals Orally Administered 5-aminolevulinic Acid for Isolation and Characterization of Circulating Tumor-Derived Extracellular Vesicles in Glioblastoma Patients

Cancers ◽  
2020 ◽  
Vol 12 (11) ◽  
pp. 3297
Author(s):  
Sybren L. N. Maas ◽  
Thomas S. van Solinge ◽  
Rosalie Schnoor ◽  
Anudeep Yekula ◽  
Joeky T. Senders ◽  
...  
Keyword(s):  
Extracellular Vesicles ◽  
Aminolevulinic Acid ◽  
Culture Media ◽  
Protoporphyrin Ix ◽  
Liquid Biopsies ◽  
Guided Surgery ◽  
Tumor Visualization ◽  
Isolation And Characterization ◽  
Droplet Pcr

Background: In glioblastoma (GB), tissue is required for accurate diagnosis and subtyping. Tissue can be obtained through resection or (stereotactic) biopsy, but these invasive procedures provide risks for patients. Extracellular vesicles (EVs) are small, cell-derived vesicles that contain miRNAs, proteins, and lipids, and possible candidates for liquid biopsies. GB-derived EVs can be found in the blood of patients, but it is difficult to distinguish them from circulating non-tumor EVs. 5-aminolevulinic acid (5-ALA) is orally administered to GB patients to facilitate tumor visualization and maximal resection, as it is metabolized to fluorescent protoporphyrin IX (PpIX) that accumulates in glioma cells. In this study, we assessed whether PpIX accumulates in GB-derived EVs and whether these EVs could be isolated and characterized to enable a liquid biopsy in GB. Methods: EVs were isolated from the conditioned media of U87 cells treated with 5-ALA by differential ultracentrifugation. Blood samples were collected and processed from healthy controls and patients undergoing 5-ALA guided surgery for GB. High-resolution flow cytometry (hFC) enabled detection and sorting of PpIX-positive EVs, which were subsequently analyzed by digital droplet PCR (ddPCR). Results: PpIX-positive EVs could be detected in conditioned cell culture media as well as in patient samples after administration of 5-ALA. By using hFC, we could sort the PpIX-positive EVs for further analysis with ddPCR, which indicated the presence of EVs and GB-associated miRNAs. Conclusion: GB-derived EVs can be isolated from the plasma of GB patients by using 5-ALA induced fluorescence. Although many challenges remain, our findings show new possibilities for the development of blood-based liquid biopsies in GB patients.

A Microfluidic Sensing System with a Multichannel Surface Plasmon Resonance Chip: Damage-free Characterization of Cells by Pattern Recognition

2020 ◽  
Author(s):  
Hiroka Sugai ◽  
Shunsuke Tomita ◽  
Sayaka Ishihara ◽  
Kyoko Yoshioka ◽  
Ryoji Kurita
Keyword(s):  
Cell Culture ◽  
Pattern Recognition ◽  
Surface Plasmon Resonance ◽  
Surface Plasmon ◽  
Plasmon Resonance ◽  
Cultured Cells ◽  
Culture Media ◽  
Biological Research ◽  
Cell Culture Media

<p>The development of a versatile sensing strategy for the damage-free characterization of cultured cells is of great importance for both fundamental biological research and industrial applications. Here, we present a pattern-recognition-based cell-sensing approach using a multichannel surface plasmon resonance (SPR) chip. The chip, in which five cysteine derivatives with different structures are immobilized on Au films, is capable of generating five unique SPR sensorgrams for the cell-secreted molecules that are contained in cell culture media. An automatic statistical program was built to acquire kinetic parameters from the SPR sensorgrams and to select optimal parameters as “pattern information” for subsequent multivariate analysis. Our system rapidly (~ 10 min) provides the complex information by merely depositing a small amount of cell culture media (~ 25 µL) onto the chip, and the amount of information obtained is comparable to that furnished by a combination of conventional laborious biochemical assays. This non-invasive pattern-recognition-based cell-sensing approach could potentially be employed as a versatile tool for characterizing cells. </p>

scholarly journals Diffusion-Based Separation of Extracellular Vesicles by Nanoporous Membrane Chip

Biosensors ◽  
2021 ◽  
Vol 11 (9) ◽  
pp. 347
Author(s):  
Gijung Kim ◽  
Min Chul Park ◽  
Seonae Jang ◽  
Daeyoung Han ◽  
Hojun Kim ◽  
...  
Keyword(s):  
Cell Culture ◽  
Human Serum ◽  
Extracellular Vesicles ◽  
Culture Media ◽  
High Yield ◽  
Polycarbonate Membrane ◽  
Cell Culture Media ◽  
Nanoporous Membrane ◽  
Selective Filter ◽  
Novel Biomarkers

Extracellular vesicles (EVs) have emerged as novel biomarkers and therapeutic material. However, the small size (~200 nm) of EVs makes efficient separation challenging. Here, a physical/chemical stress-free separation of EVs based on diffusion through a nanoporous membrane chip is presented. A polycarbonate membrane with 200 nm pores, positioned between two chambers, functions as the size-selective filter. Using the chip, EVs from cell culture media and human serum were separated. The separated EVs were analyzed by nanoparticle tracking analysis (NTA), scanning electron microscopy, and immunoblotting. The experimental results proved the selective separation of EVs in cell culture media and human serum. Moreover, the diffusion-based separation showed a high yield of EVs in human serum compared to ultracentrifuge-based separation. The EV recovery rate analyzed from NTA data was 42% for cell culture media samples. We expect the developed method to be a potential tool for EV separation for diagnosis and therapy because it does not require complicated processes such as immune, chemical reaction, and external force and is scalable by increasing the nanoporous membrane size.

A Microfluidic Sensing System with a Multichannel Surface Plasmon Resonance Chip: Damage-free Characterization of Cells by Pattern Recognition

2020 ◽  
Author(s):  
Hiroka Sugai ◽  
Shunsuke Tomita ◽  
Sayaka Ishihara ◽  
Kyoko Yoshioka ◽  
Ryoji Kurita
Keyword(s):  
Cell Culture ◽  
Pattern Recognition ◽  
Surface Plasmon Resonance ◽  
Surface Plasmon ◽  
Plasmon Resonance ◽  
Cultured Cells ◽  
Culture Media ◽  
Biological Research ◽  
Cell Culture Media

<p>The development of a versatile sensing strategy for the damage-free characterization of cultured cells is of great importance for both fundamental biological research and industrial applications. Here, we present a pattern-recognition-based cell-sensing approach using a multichannel surface plasmon resonance (SPR) chip. The chip, in which five cysteine derivatives with different structures are immobilized on Au films, is capable of generating five unique SPR sensorgrams for the cell-secreted molecules that are contained in cell culture media. An automatic statistical program was built to acquire kinetic parameters from the SPR sensorgrams and to select optimal parameters as “pattern information” for subsequent multivariate analysis. Our system rapidly (~ 10 min) provides the complex information by merely depositing a small amount of cell culture media (~ 25 µL) onto the chip, and the amount of information obtained is comparable to that furnished by a combination of conventional laborious biochemical assays. This non-invasive pattern-recognition-based cell-sensing approach could potentially be employed as a versatile tool for characterizing cells. </p>

Sign in / Sign up

Export Citation Format

Share Document