Isolation and detection of extracellular vesicles from melanoma cells and liquid biopsies using size-exclusion chromatography and nano-flow cytometry
Summary: Characterization of extracellular vesicles (EVs) holds great promise for biomarker discovery and understanding of diseases, including melanoma, the deadliest skin cancer type. Here, we describe a size-exclusion chromatography method to isolate and concentrate EVs from patient material inclu...
| Main Authors: | , , |
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| Format: | Article |
| Language: | English |
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Elsevier
2023-09-01
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| Series: | STAR Protocols |
| Subjects: | |
| Online Access: | http://www.sciencedirect.com/science/article/pii/S2666166723003325 |
| _version_ | 1797785097889382400 |
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| author | Evelyn Lattmann Valérie Lapaire Mitchell P. Levesque |
| author_facet | Evelyn Lattmann Valérie Lapaire Mitchell P. Levesque |
| author_sort | Evelyn Lattmann |
| collection | DOAJ |
| description | Summary: Characterization of extracellular vesicles (EVs) holds great promise for biomarker discovery and understanding of diseases, including melanoma, the deadliest skin cancer type. Here, we describe a size-exclusion chromatography method to isolate and concentrate EVs from patient material including (1) patient-derived melanoma cell line supernatants and (2) plasma and serum biopsies. Additionally, we provide a protocol to analyze EVs by nano-flow cytometry. EV suspensions obtained with the presented protocol can be used for several downstream analyses including RNA sequencing and proteomics. : Publisher’s note: Undertaking any experimental protocol requires adherence to local institutional guidelines for laboratory safety and ethics. |
| first_indexed | 2024-03-13T00:49:19Z |
| format | Article |
| id | doaj.art-53e79462c1634a41bc8aea94e51f8542 |
| institution | Directory Open Access Journal |
| issn | 2666-1667 |
| language | English |
| last_indexed | 2024-03-13T00:49:19Z |
| publishDate | 2023-09-01 |
| publisher | Elsevier |
| record_format | Article |
| series | STAR Protocols |
| spelling | doaj.art-53e79462c1634a41bc8aea94e51f85422023-07-08T04:19:17ZengElsevierSTAR Protocols2666-16672023-09-0143102365Isolation and detection of extracellular vesicles from melanoma cells and liquid biopsies using size-exclusion chromatography and nano-flow cytometryEvelyn Lattmann0Valérie Lapaire1Mitchell P. Levesque2Department of Dermatology, University of Zurich, University Hospital Zurich, Schlieren, ZH 8952, Switzerland; Corresponding authorDepartment of Dermatology, University of Zurich, University Hospital Zurich, Schlieren, ZH 8952, SwitzerlandDepartment of Dermatology, University of Zurich, University Hospital Zurich, Schlieren, ZH 8952, Switzerland; Corresponding authorSummary: Characterization of extracellular vesicles (EVs) holds great promise for biomarker discovery and understanding of diseases, including melanoma, the deadliest skin cancer type. Here, we describe a size-exclusion chromatography method to isolate and concentrate EVs from patient material including (1) patient-derived melanoma cell line supernatants and (2) plasma and serum biopsies. Additionally, we provide a protocol to analyze EVs by nano-flow cytometry. EV suspensions obtained with the presented protocol can be used for several downstream analyses including RNA sequencing and proteomics. : Publisher’s note: Undertaking any experimental protocol requires adherence to local institutional guidelines for laboratory safety and ethics.http://www.sciencedirect.com/science/article/pii/S2666166723003325Cell BiologyCell cultureFlow Cytometry/Mass CytometryCell separation/fractionationCancerHealth Sciences |
| spellingShingle | Evelyn Lattmann Valérie Lapaire Mitchell P. Levesque Isolation and detection of extracellular vesicles from melanoma cells and liquid biopsies using size-exclusion chromatography and nano-flow cytometry STAR Protocols Cell Biology Cell culture Flow Cytometry/Mass Cytometry Cell separation/fractionation Cancer Health Sciences |
| title | Isolation and detection of extracellular vesicles from melanoma cells and liquid biopsies using size-exclusion chromatography and nano-flow cytometry |
| title_full | Isolation and detection of extracellular vesicles from melanoma cells and liquid biopsies using size-exclusion chromatography and nano-flow cytometry |
| title_fullStr | Isolation and detection of extracellular vesicles from melanoma cells and liquid biopsies using size-exclusion chromatography and nano-flow cytometry |
| title_full_unstemmed | Isolation and detection of extracellular vesicles from melanoma cells and liquid biopsies using size-exclusion chromatography and nano-flow cytometry |
| title_short | Isolation and detection of extracellular vesicles from melanoma cells and liquid biopsies using size-exclusion chromatography and nano-flow cytometry |
| title_sort | isolation and detection of extracellular vesicles from melanoma cells and liquid biopsies using size exclusion chromatography and nano flow cytometry |
| topic | Cell Biology Cell culture Flow Cytometry/Mass Cytometry Cell separation/fractionation Cancer Health Sciences |
| url | http://www.sciencedirect.com/science/article/pii/S2666166723003325 |
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