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 transcript:

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: NIH/NINDS R01NS June 27, 2007

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

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

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

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

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

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