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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 Center
Department of Anatomy and Cell Biology
Tel: 202-994-2173
NIH/NINDS R01NS041391 June 27, 2007
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Neural Cell LineageNeural Cell Lineage
Neural Stem Cell
Progenitor Cell
Neuroblast
InhibitoryNeuron
(GABAergic)
ProjectionNeuron
(Glutamatergic)
Oligo Astrocytes
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
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Mature Neurons
Neural Stem Cells
Immature Neurons
AstrocyticCommitment
AstrocyticDifferentiation
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
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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
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CCE/GFP (Control)CCE ESCs CCE/ND6Low CCE/ND6High
MSinSB U3 R U5SD SA
CASD
U5U3
S BX
GFP
GFP
Co
un
tsF
luo
res
ce
nc
eB
rig
ht
Fie
ldA
B
U3 R U5SD SA
CASD
U5U3
S BX
ND6 GFPIRESMSin-ND6-IR-GFP-SB
MFI = 23,383 MFI = 1,148 MFI = 10,666
Generation of ND6-Expressing Mouse Embryonic Stem Cells
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NeuroD6 Promotes Neuronal Differentiation of Mouse ES Cells upon LIF and Serum Withdrawal
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Collaborators
•Dr. Robert Hawley
•Dr. Ali Ramezani
•Dr. Martine Uittenbogaard
•Teresa Hawley