APR 20, 2021 10:00 AM PDT

Use of a New 3D Pluripotent Stem Cell Suspension Culture Medium for Rapid Scale-Up and Downstream Differentiation of Pluripotent Stem Cells

Speakers
  • Scientist III, Thermo Fisher Scientific
    Biography

      Michael Akenhead is currently a Scientist III in the Biosciences Division at Thermo Fisher Scientific. In 2016, Michael received a PhD in Biomedical Engineering from the University of Kentucky. His research focused on the mechanobiology of neutrophils in the immune response. After completion of his PhD, he was hired to work at Thermo Fisher Scientific. His primary research involves working with stem cells. Specifically, Michael is exploring the ability to culture stem cells in suspension using new media formulations. The results of this research have led to the development of the Gibco™ StemScale™ PSC Suspension Medium.

    • Scientist III, Thermo Fisher Scientific
      Biography

        Michael Derr is a Scientist III in the Biosciences Division at Thermo Fisher Scientific. Michael's research at Thermo Fisher is focused on ther development and commercialization of next-generation cell culture media for pluripotent stem cell research, and media developed for neuronal differentiation and maturation applications. Michael earned his master's degree from Oregon Health and Science University (OHSU). Prior to joining Thermo Fisher Scientific in 2014,he worked in the lab of Dr. Ron Rosenfeld and Dr. Vivian Hwa at OHSU for five years where his research involved investigating the molecular signaling mechanisms of growth hormone and insulin-like growth factor 1(IGF-1) and the role they play in human growth disorders.


      Abstract

      Date:  April 20, 2021

      Time: 10:00am PDT

       

      In order to generate sufficient numbers of pluripotent stem cells (PSCs) for downstream applications, three-dimensional (3D) suspension cultures offer key advantages over two-dimensional (2D) adherent cultures. Suspension cultures are more cost-efficient and consume less plastics to grow the same number of cells in adherent cultures. However, existing solutions may be unable to fully utilize the potential of a suspension culture system. The protocols can be cumbersome and result in low fold-expansion per passage. To address these deficiencies, we recently developed a new 3D suspension culture medium – Gibco™ StemScale™ PSC Suspension Medium. StemScale medium utilizes simplified media exchanges and passaging schedules for an easy-to-use protocol. Furthermore, it enables singularized PSCs (seeded at 150,000 cells/mL) to form self-aggregating spheroids with robust expansion. Spheroids remain viable and pluripotent when grown 4 – 5 days per passage. StemScale medium maintains a consistent growth rate from small-scale orbital shaker platform cultures (well plates and shake flasks) to large-scale bioreactor cultures (3L volumes). We also demonstrate that cells obtained from dissociated spheroids can be reseeded into 3D suspension cultures, cryopreserved as single cell suspensions, or taken through downstream differentiation protocols. To demonstrate the potential of expanded PSCs in suspension with downstream applications, we have developed protocols enabling efficient and scalable neural and cardiomyocyte differentiation in 3D culture using reagents originally designed for monolayer culture. Key parameters and considerations have been identified and optimized for suspension culture differentiation from StemScale PSC spheroids. This approach has yielded wide-ranging benefits including faster differentiation and expansion steps, scalable expansion of neural stem cells (NSCs) and neural progenitor cells, and earlier onset of neuronal maturation and functional activity.

       

      Learning Objectives:

      • Discuss and learn how to achieve a high pluripotent stem cells (PSCs) yield (5 – 10 fold expansion per passage) in suspension culture.
      • Describe and understand how to optimize PSC spheroid growth in various suspension culture vessels and bioreactors.
      • Discuss and learn how to prepare and use 3D spheroids in downstream differentiation applications.

       

       

       

      Webinars will be available for unlimited on-demand viewing after live event.


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