Efficient generation of midbrain and hindbrain neurons from mouse embryonic stem cells.
Embryonic stem (ES) cells are clonal cell lines derived from the inner cell mass of the developing blastocyst that can proliferate extensively in vitro and are capable of adopting all the cell fates in a developing embryo. Clinical interest in the use of ES cells has been stimulated by studies showing that isolated human cells with ES properties from the inner cell mass or developing germ cells can provide a source of somatic precursors. Previous studies have defined in vitro conditions for promoting the development of specific somatic fates, specifically, hematopoietic, mesodermal, and neurectodermal. In this study, we present a method for obtaining dopaminergic (DA) and serotonergic neurons in high yield from mouse ES cells in vitro. Furthermore, we demonstrate that the ES cells can be obtained in unlimited numbers and that these neuron types are generated efficiently. We generated CNS progenitor populations from ES cells, expanded these cells and promoted their differentiation into dopaminergic and serotonergic neurons in the presence of mitogen and specific signaling molecules. The differentiation and maturation of neuronal cells was completed after mitogen withdrawal from the growth medium. This experimental system provides a powerful tool for analyzing the molecular mechanisms controlling the functions of these neurons in vitro and in vivo, and potentially for understanding and treating neurodegenerative and psychiatric diseases.
Lee
S H
SH
Laboratory of Molecular Biology, NINDS, NIH, Bethesda, MD 20892, USA.
Lumelsky
N
N
Studer
L
L
Auerbach
J M
JM
McKay
R D
RD
D000818:Animals; D001205:Ascorbic Acid / Q000494:pharmacology; D018929:Cell Culture Techniques / Q000379:methods; D002454:Cell Differentiation; D002455:Cell Division; D002460:Cell Line; D002851:Chromatography, High Pressure Liquid; D000242:Cyclic AMP / Q000494:pharmacology; D004298:Dopamine / Q000378:metabolism; D004622:Embryo, Mammalian; D016222:Fibroblast Growth Factor 2 / Q000378:metabolism; D051524:Fibroblast Growth Factor 8; D005346:Fibroblast Growth Factors / Q000494:pharmacology; D018698:Glutamic Acid / Q000494:pharmacology; D053823:Hedgehog Proteins; D007150:Immunohistochemistry; D008636:Mesencephalon / Q000166:cytology*; D051379:Mice; D009474:Neurons / Q000166:cytology*; D018408:Patch-Clamp Techniques; D011506:Proteins / Q000494:pharmacology; D020133:Reverse Transcriptase Polymerase Chain Reaction; D012249:Rhombencephalon / Q000166:cytology*; D012701:Serotonin / Q000378:metabolism; D013234:Stem Cells / Q000166:cytology*; D013779:Tetrodotoxin / Q000494:pharmacology; D015534:Trans-Activators; D005680:gamma-Aminobutyric Acid / Q000494:pharmacology
C496457:Fgf8 protein, mouse; D053823:Hedgehog Proteins; D011506:Proteins; D015534:Trans-Activators; D016222:Fibroblast Growth Factor 2; D051524:Fibroblast Growth Factor 8; D012701:Serotonin; D018698:Glutamic Acid; D013779:Tetrodotoxin; D005680:gamma-Aminobutyric Acid; D005346:Fibroblast Growth Factors; D000242:Cyclic AMP; D001205:Ascorbic Acid; D004298:Dopamine