NOV 17, 2016 07:00 AM PST

From heart to mind: Simplified iPS cell reprogramming into cardiac and neural cells under hypoxic conditions

SPONSORED BY: Eppendorf, Eppendorf
C.E. CREDITS: P.A.C.E. CE | Florida CE
  • Research Associate Director, Stem Cell Training Course; Rutgers University

      Rick I Cohen is the Director of the Stem Cell Training Course and Research Associate in the Department of Biomedical Engineering at Rutgers University in Piscataway, NJ. Over the past 8+ years he has been instrumental in providing cell based materials and hands-on training to participants from both academia and industry. In the past, he directed the Rutgers Stem Cell Core Facility, and has trained the first generation of T32 multi-institutional stem cell trainees. He has worked with Peprotech in Rocky Hill, NJ to develop several commercialized stem cell media and related products. He plans to continue his efforts to offer engineered cellular solutions to answer questions about cellular reprogramming and differentiation to needed cell types.

      One of the on-going projects in Dr. Cohen's lab examines if Dopaminergic Neuron and Cardiogenic differentiation of IPSCs be optimized using small molecules. For the dopaminergic differentiation project, his lab is collaborating with Jared Sterneckert from the Center for Regenerative Therapies in Dresden, Germany and PeproTech to determine the efficacy of a two step protocol to generate floor plate neuro-epithelia and then direct these cells into dopaminergic neurons as stimulated by small molecules or growth factors in low oxygen tension. The aim of this study is to determine the optimized and scalable method to produce functional dopaminergic neurons for studies in Parkinson's Disease. For cardiomyocyte differentiation, his lab is reproducing and harmonizing recent publications using a simplified serial combination of small molecules to inhibit GSK3-beta pathway followed by inhibition of the Wnt signaling pathway. The final focus of this project is to seed the cardiomyocytes or their progenitors on a bio-compatible and transplantable scaffold in order to create a cardiac patch capable of restoring function to damaged myocardium.

    DATE: November 17, 2016
    TIME: 7:00am PT, 10:00am ET

    Induced pluripotent stem (iPS) cell reprogramming allows turning an adult somatic cell into a pluripotent stem cell. Four Factor pluripotency reprogramming of somatic cells and subsequent multi-lineage directed differentiation can be accomplished by robust genetically modifying/manipulating methods. More recently non-genetically modifying methods for induction have gained popularity as they lend themselves to translational and clinically related research paradigms. 

    In this study we developed an optimized single Episomal vector bearing multiple interspersed genes. This single entity plasmid electroporated into fibroblasts and cultured in low oxygen tension (4%O2) with optimized xeno-free media and accompanying small molecule mixtures leads to reproducible reprogramming. Simplifications of xeno-free neuronal and cardiac differentiation methods rely on a 2 step procedure. Differentiated neuronal cells become TH+/MAP2+, and secrete dopamine as measured by HPLC and differentiated cardiac cell leads to efficient appearance of beating clusters within 8-10 days of initiating the protocol. In order to ready methods for translational and clinical type research, the goal of the study presented here was to optimize the culturing atmosphere, and simplify the regimen of medias, growth factors, and small molecules that will streamline multiple differentiation pathways.

    Learning Objectives:
    • Obtain an overview of somatic cell reprogramming methods
    • Learn about episomal repogramming methodology and its advantages
    • Learn how to differentiate induced pluripotent cells into neurons and functional cardiomyocytes
    • Gain insights into improving efficiency of cellular reprogramming by optimizing culture atmosphere, media, growth factors and small molecules

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