Differentiation of human dopamine neurons from an embryonic carcinomal stem cell line.
Previous studies from this laboratory have demonstrated that fibroblast growth factor 1 together with a number of co-activator molecules (dopamine, TPA, IBMX/forskolin), will induce the expression of the catecholamine biosynthetic enzyme tyrosine hydroxylase (TH) in 10% of human neurons (hNTs) derived from the NT2 cell line [10]. In the present study, we found that TH induction was increased to nearly 75% in hNTs when cells were permitted to age 2 weeks in culture prior to treatment with the differentiation cocktail. This high level of TH expression was sustained 7 days after removal of the differentiating agents from the media. Moreover, the induced TH present in these cells was enzymatically active, resulting in the production of low levels of dopamine (DA) and its metabolite DOPAC. These findings suggest that hNTs may provide an important tissue culture model for the study of factors regulating TH gene expression in human neurons. Moreover, hNTs may serve, in vivo, as a source of human DA neurons for use in transplantation therapies.
Iacovitti
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Department of Neurology, Thomas Jefferson University Medical College, 1025 Walnut Street, Philadelphia, PA 19107, USA. lorraine.iacovitti@mail.tju.edu
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D015056:1-Methyl-3-isobutylxanthine / Q000494:pharmacology; D015102:3,4-Dihydroxyphenylacetic Acid / Q000378:metabolism; D002454:Cell Differentiation / Q000187:drug effects* / Q000502:physiology; D005576:Colforsin / Q000494:pharmacology; D046508:Culture Techniques; D004298:Dopamine / Q000096:biosynthesis* / Q000494:pharmacology; D016220:Fibroblast Growth Factor 1 / Q000494:pharmacology; D006801:Humans; D007150:Immunohistochemistry; D008954:Models, Biological; D014411:Neoplastic Stem Cells / Q000166:cytology / Q000187:drug effects* / Q000201:enzymology; D009474:Neurons / Q000166:cytology / Q000187:drug effects* / Q000201:enzymology; D010300:Parkinson Disease / Q000378:metabolism / Q000503:physiopathology; D010726:Phosphodiesterase Inhibitors / Q000494:pharmacology; D010959:Tissue Plasminogen Activator / Q000494:pharmacology; D014446:Tyrosine 3-Monooxygenase / Q000187:drug effects* / Q000378:metabolism
D010726:Phosphodiesterase Inhibitors; D015102:3,4-Dihydroxyphenylacetic Acid; D016220:Fibroblast Growth Factor 1; D005576:Colforsin; D014446:Tyrosine 3-Monooxygenase; D010959:Tissue Plasminogen Activator; D015056:1-Methyl-3-isobutylxanthine; D004298:Dopamine