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Genetic Manipulation of Mouse ES Cells toward a Specific Neuronal Fate Anne Chiaramello, Ph.D Associate Professor George Washington University Medical.

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Presentation on theme: "Genetic Manipulation of Mouse ES Cells toward a Specific Neuronal Fate Anne Chiaramello, Ph.D Associate Professor George Washington University Medical."— Presentation transcript:

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 NIH/NINDS R01NS041391 June 27, 2007

2 Neural Cell Lineage Neural Stem Cell Progenitor Cell Neuroblast Inhibitory Neuron (GABAergic) Projection Neuron (Glutamatergic) OligoAstrocytes Glioblast LIF ? 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 bFGF

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

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

5 CCE/GFP (Control)CCE ESCsCCE/ND6 Low CCE/ND6 High MSinSB U3RU5 SDSA CA SD U5  U3 S B X GFP Counts Fluorescence Bright Field A B U3RU5 SDSA CA SD U5  U3 S B X ND6 GFP IRES MSin-ND6-IR-GFP-SB MFI = 23,383MFI = 1,148MFI = 10,666 Generation of ND6-Expressing Mouse Embryonic Stem Cells

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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