Production of tyrosine hydroxylase positive neurons
AssigneeStem Cell Therapeutics Inc.
InventorsSamuel Weiss, Tetsuro Shingo
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Abstract
The present invention relates to a method of producing neurons that express the enzyme tyrosine hydroxylase (TH) by subjecting neural stem cells to FGF-1, a protein kinase A activator, a protein kinase C activator, and dopamine/L-DOPA. Surprisingly, when forskolin is used as a protein kinase A activator, it requires only low levels of FGF-1 and forskolin to efficiently produce TH positive neurons from fetal or adult neural stem cells. Also provided are compositions used to produce TH positive neurons and the resulting neural cell culture, as well as a method of treating disease or conditions which are associated with dopamine neuron loss or dysfunction.
Claims38
A composition useful for producing TH positive neurons from neural stem cells, comprising fibroblast growth factor 1 (FGF-1), a protein kinase A activator selected from the group consisting of isobutylmethylxanthine (IBMX), pituitary adenylate cyclase activation polypeptide (PACAP), forskolin, and any combination thereof, a phorbol ester, and dopamine/L-DOPA, with the proviso that if the protein kinase A activator is a combination of isobutylmethylxanthine (IBMX) and forskolin, the concentration of forskolin is less than about 3 μM.
A pharmaceutical composition comprising fibroblast growth factor 1 (FGF-1), a protein kinase A activator selected from the group consisting of isobutylmethylxanthine (IBMX), pituitary adenylate cyclase activating polypeptide (PACAP), forskolin, and any combination thereof;
- a phorbol ester, dopamine/L-DOPA, and one or more selected from the group consisting of epidermal growth factor (EGF), bone morphogenic protein 2 (BMP-2) and brain derived neurotrophic factor;
- and a pharmaceutically acceptable excipient and/or carrier with the proviso that if the protein kinase A activator is a combination of isobutylmethylxanthine (IBMX) and forskolin, the concentration of forskolin is less than about 3 μM.
Dependencies are inferred from the claim text.