NOV 07, 2018 7:00 AM PST

Analysis of Surface Antigens on Exosomes

Speaker
  • Manager, Basic & Translational Research Operations, Chief, Flow Cytometry Core Laboratory, Mitchell Cancer Institute, University of South Alabama, United States
    Biography
      Steve McClellan, BS, MT, SCYM (ASCP)CM is a life science professional with over 30 years of expertise in advanced flow cytometry & cell sorting. He has worked in basic and clinical research in the areas of cancer biology, stem cell therapy, transplant immunology and xenotransplantation. He is currently Manager of Basic & Translational Research Operations at the University of South Alabama Mitchell Cancer Institute, where he also serves as Chief, Flow Cytometry Core Laboratory. For the past 12 years, his lab has been conducting research on cancer stem cells (CSC) and circulating tumor cells (CTC); working to develop better methods of purification, culture and analysis at both genetic and functional levels, as well as using CSC in HTS drug discovery. Exosomes are the latest addition to the lab's research efforts, working primarily with patient samples to measure response to therapy and to look for early diagnostic biomarkers. His specialty interests include: Biomarkers, Cancer Research, Cancer Therapeutics, Cell Signalling/Transduction, Clinical Research, High Throughput Screening, Immunology, Immuno-Oncology, Oncology, Stem Cell Technologies,.

    Abstract

    DATE: November 7, 2018
    TIME: 7:00AM PDT

    Exosomes have been shown to have significant roles in cancer including disease progression acting in the tumor micro-environment, metastasis acting in the peripheral circulation and immune suppression acting in both areas. We have sought to translate information gathered from exosomes isolated from cancer cell culture supernatant, to those isolated from pancreatic cancer plasma and cyst fluid obtained by endoscopy. Biomarker discovery is of the utmost importance for this deadly form of cancer. Traditional flow cytometers generally struggle to resolve individual exosomes.  We have previously demonstrated that the use of ultra-filtered sheath fluid (0.025 um pore size) significantly increases the resolution of exosomes. We use the Invitrogen Attune NxT Flow Cytometer to analyze individual exosomes for the expression of a variety of surface markers using multi-color staining.

    Learning objectives:

    • Different exosome isolation methods 
    • Best practices for the flow cytometric analysis of exosomes 
    • Surface marker analysis of exosomes

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