November 17, 2016
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.
- 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