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Flow Cytometric Analysis of Extracellular Vesicles from Cell-conditioned Media
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Flow Cytometric Analysis of Extracellular Vesicles from Cell-conditioned Media

Flow Cytometric Analysis of Extracellular Vesicles from Cell-conditioned Media

DOI:
10.3791/59128-v

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08:32 min

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February 12, 2019

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, , ,
1Cellular and Molecular Cardiology Laboratory,Cardiocentro Ticino Foundation, Switzerland, 2Molecular Cardiology Institute, Dept. of Cardiology,University of Zürich, 3Faculty of Biomedical Science,Università Svizzera Italiana, Switzerland

Chapters

  • 00:04Title
  • 01:05Conditioned Medium Collection and Processing
  • 02:45Nanoparticle Tracking Analysis (NTA) Quantification
  • 03:27Sample Preparation
  • 04:17Sample Acquisition and Analysis
  • 06:32Results: Representative Flow Cytometric Analyses of Extracellular Vesicles from Cell-conditioned Medium
  • 07:58Conclusion

Summary

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The protocol describes a reproducible method designed for use with cell culture supernatants to detect surface epitopes on small extracellular vesicles (EV). It utilizes specific EV immunoprecipitation using beads coupled with antibodies that recognize surface antigen CD9, CD63, and CD81. The method is optimized for downstream flow cytometry analysis.

Tags

Flow CytometryExtracellular VesiclesCell-conditioned MediaBiomarkersTherapeutic ToolsDiagnosticParticle CharacterizationSurface MarkersCardiac Progenitor CellsExosome ProductionUltracentrifugationNanoparticle Tracking Analysis

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