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Selection and Extraction of Promoter-Defined Neural Stem Cells from Mixed Cultures of Human Embryonic Stem Cells

*Abstract: This invention describes the use of promoter-based GFP selection technology to identify and isolate neuronal progenitor cells, oligodendrocyte progenitor cells, or neural stem cells from mixed populations of human embryonic stem cells. The isolated neural stem cells are multipotent, can be maintained in continuously expanding culture, and can generate both neurons and glia. These embryonic neural cells engraft well upon transplantation, and can effectively integrate as neurons, oligodendrocytes or astrocytes, thereby restoring cells lost to brain disease and injury.

Neuronal progenitor cells may be separated from the mixed population using a ta1 tubulin promoter, a Hu promoter, an ELAV promoter, a MAP-1B promoter, or a GAP-43 promoter.

Oligodendrocyte progenitor cells may be separated from the mixed population using a CNP promoter, an NCAM promoter, a myelin basic protein promoter, a JC virus minimal core promoter, a myelin-associated glycoprotein promoter, or a proteolipid protein promoter.

Neural stem cells may be separated from the mixed population using a musashi promoter or the nestin enhancer.

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