Rapid and efficient differentiation of dopaminergic neurons from mouse embryonic stem cells.
We have developed a fast and effective method for the differentiation of dopaminergic neurons from mouse embryonic stem cells. Neuronal precursors are obtained by formation of embryonic bodies or neural stem spheres via free-floating culture in the presence of the mitogens basic fibroblast growth factor and epidermal growth factor together with L-ascorbic acid. Subsequent culturing of the precursor cells in medium containing epidermal growth factor, FGF8b, SHH and ascorbic acid induces cell proliferation, following withdrawal of the growth factors leads differentiation into predominantly dopaminergic neurons. Mature neurons are obtained within 10 days of replacing the proliferation to differentiation medium. Embryonic stem-derived dopaminergic neurons are purified by cell sorting and may serve as a convenient source for the study of molecular, genetic and cellular properties of dopaminergic neurons.
Lau
Thorsten
T
Biochemical Laboratory, Central Institute of Mental Health, Mannheim, Germany.
Adam
Sylvia
S
Schloss
Patrick
P
D000818:Animals; D002454:Cell Differentiation / Q000187:drug effects / Q000502:physiology*; D049109:Cell Proliferation / Q000187:drug effects; D002478:Cells, Cultured; D017077:Culture Media, Conditioned / Q000494:pharmacology; D004298:Dopamine / Q000378:metabolism*; D004622:Embryo, Mammalian; D005434:Flow Cytometry / Q000379:methods; D006133:Growth Substances / Q000494:pharmacology; D007150:Immunohistochemistry / Q000379:methods; D051379:Mice; D009419:Nerve Tissue Proteins / Q000378:metabolism; D009474:Neurons / Q000187:drug effects / Q000502:physiology*; D013234:Stem Cells / Q000187:drug effects / Q000502:physiology*
D017077:Culture Media, Conditioned; D006133:Growth Substances; D009419:Nerve Tissue Proteins; D004298:Dopamine