genetic manipulation of mouse es cells toward a specific neuronal fate anne chiaramello, ph.d...
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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
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
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
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
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
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