MAR 24, 2020 11:00 AM PDT
Add to Calendar Select one of the following: iCal Google Calendar Outlook Calendar Yahoo Calendar

3-D Organoid Models for Differentiation of PSC-Derived Dopaminergic Neurons

Speakers
  • Sr. Staff Scientist, Cell Biology 3D Models, Thermo Fisher Scientific
    Biography
      Richard Josephson earned his Ph.D. in Cell and Molecular Biology at the Massachusetts Institute of Technology. He conducted post-doctoral research on gene regulation by stem cells of the central nervous system under Ron McKay in the National Institute of Neurological Disorders and Stroke at NIH. He was subsequently a research scientist in the Stem Cell Center at the American Type Culture Collection, where he was lead author of several papers and reviews on authentication and standards for human embryonic stem cells and stem cell therapies. Later, as a Senior Scientist with MTI-GlobalStem, he developed products and methods for neurobiology including large-scale production and long-term culture of rodent primary neurons and human iPSC-derived neurons. At Thermo Fisher Scientific, Dr. Josephson is involved in the development and analysis of new 3-D cell culture models including midbrain organoids to study Parkinson Disease and other neurological disorders.

    Abstract:
    DATE:   March 24, 2020
    TIME:   11:00am PT, 2:00pm ET
     
     
    While in vitro cell culture has long been used to study neurological diseases, researchers have come to the realization that 2-D systems do not accurately model the complex cell interactions and self-organizing architecture of the brain. Neuroscientists are increasingly turning to 3-D organoid systems that include a greater diversity of cell types and more closely mimic brain development in vivo.
     
    We have generated organoids rich in midbrain dopaminergic (DA) neurons, the specific cells which degenerate in Parkinson's Disease (PD). Starting with suspension culture of pluripotent stem cells, we passage into low-attachment 96-well U-bottom plates to begin differentiation of uniformly-sized spheroids. A low concentration of extracellular matrix (ECM) molecules are added during neural specification to enhance the organoid architecture, while improving throughput relative to ECM encapsulation methods. The ECM speeds dopaminergic maturation as indicated by Neuromelanin production. Multielectrode array (MEA) recording of replated spheroids shows spontaneous burst activity in as little as five weeks, followed by gradual refinement toward coordinated rhythmic bursting. To model Parkinson's Disease in vitro we introduced the PD-related SNCA A30P mutation into a Cas9-expressing iPSC line. We have generated midbrain organoids from these lines and are evaluating the sensitivity of mutant and wildtype DA neurons to induced oxidative stress. The specific effects on DA neurons are assayed by antibody detection of active caspase in wholemount or cryosectioned organoids and evaluated via High-Content Analysis (HCA).
     
    In this work we have produced midbrain organoids with functional DA neurons while reducing our time and effort. In combination with more accessible and higher-throughput 3-D assays, we hope this may contribute toward a more illuminating in vitro PD model.
     
    Learning Objectives:
    • Discover the advantages of 3-D culture for directed neuronal differentiation of PSC
    • Learn how to modify early specification steps to expedite neuronal maturation
    • Understand the constraints that must be balanaced to adapt 2-D differentiation protocols to 3-D
     
     
     
    Webinars will be available for unlimited on-demand viewing after live event.
     
    LabRoots is approved as a provider of continuing education programs in the clinical laboratory sciences by the ASCLS P.A.C.E. ® Program. By attending this webinar, you can earn 1 Continuing Education credit once you have viewed the webinar in its entirety.
     
     

    Show Resources
    You May Also Like
    JAN 23, 2020 9:00 AM PST
    C.E. CREDITS
    JAN 23, 2020 9:00 AM PST
    DATE: January 23, 2020 TIME: 9:00am PST, 12:00pm EST...
    NOV 18, 2019 7:00 AM PST
    C.E. CREDITS
    NOV 18, 2019 7:00 AM PST
    DATE: November 18, 2019TIME: 7:00am PST, 11:00am EST, 4:00pm CEWT How often do you pipette in your cell culture lab every day? Usually, we do it so often that we tend stop th...
    SEP 05, 2019 4:00 PM CEST
    C.E. CREDITS
    SEP 05, 2019 4:00 PM CEST
    DATE: September 5, 2019TIME: 7:00am PT, 10:00am ET, 4:00pm CEST PCR (Polymerase Chain Reaction) has gone through a massive evolution since its development in 1983. Besides it...
    OCT 02, 2019 11:00 AM PDT
    OCT 02, 2019 11:00 AM PDT
    DATE: October 2, 2019TIME: 11:00am PDT, 2:00pm EDT Ditch the Excel spreadsheets and manage your molecular workflows entirely in your LIMS Achieve configuration of molecular workf...
    FEB 26, 2020 9:00 AM PST
    C.E. CREDITS
    FEB 26, 2020 9:00 AM PST
    DATE: February 26, 2020 TIME: 9:00am PST 3D cell culture and analysis and the study of organoids and spheroids are becoming more prevalent as a research method in publications as traditional...
    MAR 24, 2020 10:00 AM PDT
    Add to Calendar Select one of the following: iCal Google Calendar Outlook Calendar Yahoo Calendar
    C.E. CREDITS
    MAR 24, 2020 10:00 AM PDT
    Add to Calendar Select one of the following: iCal Google Calendar Outlook Calendar Yahoo Calendar
    DATE: March 24, 2020 TIME: 10:00 am PDT, 1:00pm EDT The Clampfit software module is a useful tool to manipulate and analyze electrophysiological data acquired by pCLAMP™ software. Rece...
    Loading Comments...
    Show Resources
    Event Countdown
    • 0 Days
    • 0 Hours
    • 0 Minutes
    • 0 Seconds