SLAS2020 Short Courses

Set-up and Validation of 3D Primary, Stem Cell and Immortalized Cultures for Downstream Microplate Reader and Imaging Applications

Mammalian cell culture has long been an invaluable tool in cell biology, drug discovery and regenerative medicine. When cell culture techniques were first developed, 3-Dimensional (3D) systems were utilized. That rapidly changed due to cost and efficiency concerns resulting in cell culture now being performed with adherent cells grown on flat and rigid two-dimensional (2D) substrates including polystyrene and glass. Advances in our understanding of cell physiology and failures in clinical trials have provided the impetus to move away from 2D systems and back to a more in-vivo-like 3D environment. The advances of these new technologies and screening methodologies have allowed scientists to assess more realistic functional capabilities of cells. The following course will focus on the cellular microenvironment and its importance when developing and screening cell-based assays using primary, stem cell and immortalized cultures in 3D systems.

Who Should Attend

• Industry and academic scientists
• Those interested or currently involved in 3D culture systems
• Those with mid to advanced-level experience in cell-based assays or liquid handling, microplate reading and imaging technologies

Course Benefits:

• Learn helpful hints for setting up, validating and assessing a variety of cell types in a 3D environment
• Utilize automation in setting up, validating and screening 3D cultures
• Participate in a focused discussion on assay types and microplate reading and imaging techniques that are currently in use for screening in 3D

Course Topics:

• Introduction
  • Understanding the cell-based assay microenvironment
    • Importance of the chosen platform
    • Medium considerations
    • Surface and ECM’s
  • 2-Dimensional vs 3-Dimensional cell culture: Compare and Contrast
  • Summary
• Setting up and validating 3D systems
  • Optimizing the 3D microenvironment
    • Media
    • ECM’s
    • Cells
      • Tumor Spheroids
      • Stem cells
      • Organoids
  • Automating the process
    • Liquid handling
      • 3D cell models in ECM or free floating
      • How to perform media exchanges or washes without disturbing cellular structures
    • Microplate/system handling
      • Large integrated system vs. smaller, modular instrumentation
      • Containment, aseptic processing needs
  • An introduction to assaying in 3D
    • What techniques are available to screen 3D systems
      • Microplate based readers
        • Systems used in 2D and 3D
        • Assaying Kits
          • Traditional kits for assaying 2D
          • Advanced methods; 2D kits being used for 3D applications
          • Kits developed for 3D
      • Imaging
        • Systems used in 2D and 3D
        • Assaying Kits
          • Traditional techniques for assaying 2D
          • Advanced methods; 2D kits being used for 3D applications
          • Kits developed for 3D
    • Considerations when setting up 3D assays
      • Reading and imaging using current 3D technologies
      • Setting up the assay using LH and automation in a 3D environment
  • Summary
  • Case studies from the field, use of Field Application Scientists to relate field experiences.

Instructors:

Austin Mogen, Ph.D. Corning Austin Mogen, Ph.D., is a Field Application Scientist at Corning Life Sciences. He received his doctorate from the University of Florida before working as a Senior Scientist of upstream process development and supervisor for manufacturing of viral vectors. In these positions he focused on bioprocess development, closed system solutions for cell culture scale-up and production of viral vectors for cell and gene therapy. Mogen works extensively with academic researchers and process development groups, optimizing cell culture assays and cellular scale-up conditions. In addition, he focuses on collaborations utilizing high-end 3D technology products such as the Corning spheroid microplates, transwell inserts and extracellular matrices to provide more in vivo models for biological research and therapeutic response.

Joe Clayton, Ph.D. BioTek Joe Clayton, Ph.D., is a Principal Scientist at BioTek Instruments, Inc. He received his Ph.D. in Molecular, Cell and Biomedical Sciences from the University of Vermont and his M.S. in Healthcare Research from The Dartmouth Institute. Clayton’s primary role at BioTek is developing and optimizing kinetic live cell imaging applications, utilizing advanced 2D and 3D systems.