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Genetic Manipulation of Mouse ES Cells toward a Specific Neuronal Fate

Genetic Manipulation of Mouse ES Cells toward a Specific Neuronal Fate. Anne Chiaramello, Ph.D Associate Professor George Washington University Medical Center Department of Anatomy and Cell Biology Tel: 202-994-2173 anaaec@gwumc.edu. June 27, 2007. NIH/NINDS R01NS041391. Objective.

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Genetic Manipulation of Mouse ES Cells toward a Specific Neuronal Fate

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  1. Genetic Manipulation of Mouse ES Cells toward a Specific Neuronal Fate Anne Chiaramello, Ph.D Associate Professor George Washington University Medical Center Department of Anatomy and Cell Biology Tel: 202-994-2173 anaaec@gwumc.edu June 27, 2007 NIH/NINDS R01NS041391

  2. Objective To direct neural fate of mouse ES Cells by genetic manipulation for cell replacement therapies in the context of specific neurodegenerative disorders and CNS injuries. Embryonic Stem Cell LIF Neural Cell Lineage bFGF Neural Stem Cell Progenitor Cell Glioblast ? Neuroblast Inhibitory Neuron (GABAergic) Projection Neuron (Glutamatergic) Oligo Astrocytes

  3. Strategy to Direct Fate of Mouse ES Cells NeuroD6 Ngn1/2 Initiation of Differentiation Cell Cycle Exit Subtype Specification Neural Commitment Terminal Differentiation NeuralProgenitor Immature Neurons Neural Stem Cells Mature Neurons Astrocytic Commitment AstrocyteProgenitor Astrocytes Astrocytic Differentiation

  4. Committed Neural Progenitor Cells (proliferative & post-mitotic) Neural Stem Cells Glutamatergic Pyramidal Neurons (Upper Motor Neurons) Wu, Sheng-Xi et al. (2005) Proc. Natl. Acad. Sci. USA 102, 17172-17177 PC12-NeuroD6 Cells NeuroD6-Mediated Glutamatergic Pyramidal Fate and Corticogenesis

  5. X MSinSB U5 B CA S GFP SD SD SA U3 U3 U3 R R U5 U5 X MSin-ND6-IR-GFP-SB U5 CA GFP S ND6 IRES B SD SD SA U3 CCE ESCs CCE/GFP (Control) CCE/ND6Low CCE/ND6High Counts GFP Generation of ND6-Expressing Mouse Embryonic Stem Cells A B MFI = 23,383 MFI = 1,148 MFI = 10,666 Fluorescence Bright Field

  6. NeuroD6 Promotes Neuronal Differentiation of Mouse ES Cells upon LIF and Serum Withdrawal

  7. Collaborators • Dr. Robert Hawley • Dr. Ali Ramezani • Dr. Martine Uittenbogaard • Teresa Hawley

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