Generation of cortical neurons from mouse embryonic stem cells.
Embryonic stem cells (ESCs) constitute a tool of great potential in neurobiology, enabling the directed differentiation of specific neural cell types. We have shown recently that neurons of the cerebral cortex can be generated from mouse ESCs cultured in a chemically defined medium that contains no morphogen, but in the presence of the sonic hedgehog inhibitor cyclopamine. Corticogenesis from ESCs recapitulates the most important steps of cortical development, leading to the generation of multipotent cortical progenitors that sequentially produce cortical pyramidal neurons displaying distinct layer-specific identities. The protocol provides a most reductionist cellular model to tackle the complex mechanisms of cortical development and function, thereby opening new perspectives for the modeling of cortical diseases and the design of novel neurological treatments, while offering an alternative to animal use. In this protocol, we describe a method by which millions of cortical neurons can be generated in 2-3 weeks, starting from a single frozen vial of ESCs.
Gaspard
Nicolas
N
Institute of Interdisciplinary Research on Human and Molecular Biology (IRIBHM), Université Libre de Bruxelles (ULB), Brussels, Belgium.
Bouschet
Tristan
T
Herpoel
Adèle
A
Naeije
Gilles
G
van den Ameele
Jelle
J
Vanderhaeghen
Pierre
P
D000818:Animals; D018929:Cell Culture Techniques; D002454:Cell Differentiation; D002540:Cerebral Cortex / Q000166:cytology*; D003470:Culture Media; D053595:Embryonic Stem Cells / Q000166:cytology*; D005455:Fluorescent Antibody Technique; D051379:Mice; D009474:Neurons / Q000166:cytology*; D020133:Reverse Transcriptase Polymerase Chain Reaction
D003470:Culture Media