Production of midbrain dopaminergic neurons and methods for the use thereof
AssigneeCellular Dynamics International, Inc.
InventorsMatt GEORGE, Carrie CHAVEZ, Chris McMAHON, Wen Bo WANG, Lucas CHASE, Brad SWANSON
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Abstract
Methods are provided for efficient production of midbrain dopaminergic (DA) neurons. In some aspects, methods involve differentiation and selection of DA neurons for a transgenic pluripotent cell population (e.g., cells comprising a selectable marker gene). Cell populations produced by the instant methods and methods of their use are likewise provided.
Claims17
1-44. (canceled)
A method for providing an enriched population of midbrain dopaminergic (DA) neurons comprising:
- differentiating cells of a population of human induced pluripotent cells to provide a neural lineage cell population;
- further differentiating cells of the neural lineage cell population to generate a cell population which includes midbrain neurons;
- and
- purifying cells from said cell population using a transgenic screenable or selectable marker under the control of a pan-neural promoter expressed by cells of the cell population, to provide an enriched population of midbrain DA neurons;
- wherein the differentiation differentiation steps are carried out in a chemically defined medium that is free of feeder cells and feeder cell extracts and at least 80% of cells in the enriched population are positive for both Lmx1 and FoxA2 expression.
A method for providing an enriched population of midbrain dopaminergic (DA) neurons comprising:
- obtaining a population of human induced pluripotent cells;
- differentiating the cells into a neural lineage cell population in a medium comprising a MEK inhibitor;
- and
- further differentiating cells of the neural lineage cell population to provide an enriched population of midbrain DA neurons;
- and
- wherein said differentiation steps are carried out in chemically defined medium that is free of feeder cells and feeder cell extracts and at least 80% of cells in the enriched population are positive for both Lmx1 and FoxA2 expression.
Dependencies are inferred from the claim text.