METHODS OF GENERATING EPITHELIAL LINEAGE CELLS FROM EMBRYOID BODIES AND PLURIPOTENT CELLS
First Claim
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1. A method of generating p63-positive cells, the method comprising the step of:
- culturing embryoid bodies in a medium comprising a retinoid and optionally a bone morphogenetic protein for about two days to about nine days, each in an amount sufficient to generate p63-positive cells.
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Abstract
Methods of generating p63-positive cells from embryoid bodies and pluripotent cells by culturing the cells in the presence of a retinoid and optionally a bone morphogenetic protein, such that the cells express at least p63. The p63-positive cells can be further cultured without the retinoid and optional bone morphogenetic protein to K14-positive cells. The K14-positive cells can be further cultured into various terminally differentiated cell types of the epithelial lineage.
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Citations
27 Claims
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1. A method of generating p63-positive cells, the method comprising the step of:
culturing embryoid bodies in a medium comprising a retinoid and optionally a bone morphogenetic protein for about two days to about nine days, each in an amount sufficient to generate p63-positive cells. - View Dependent Claims (2, 3, 4, 5, 6, 7, 8)
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9. A method of generating p63 positive cells, the method comprising the step of:
culturing pluripotent cells in a medium comprising a retinoid and optionally a bone morphogenetic protein for about five days to about seven days, each in an amount sufficient to generate p63 positive cells. - View Dependent Claims (10, 11, 12, 13, 14, 15, 16)
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17. A method of generating K14-positive cells, the method comprising the step of:
culturing p63-positive cells to confluence in a defined serum-free medium to obtain K14-positive cells, wherein the cells are on an adherent surface, and wherein the medium does not comprise a retinoid and bone morphogenetic protein. - View Dependent Claims (18)
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19. A method of generating filaggrin- and involucrin-positive cells, the method comprising the step of:
culturing K14-positive cells to confluence in a defined serum-free medium to obtain filaggrin- and involucrin-positive cells, wherein the cells are on an adherent surface. - View Dependent Claims (20, 21)
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22. A method of generating K3/K12-positive cells, the method comprising the step of:
culturing K14-positive cells to confluence in a defined serum-free medium to obtain K3/K12-positive cells, wherein the cells are on an adherent surface. - View Dependent Claims (23)
- 24. A population of cultured K14-positive cells, wherein at least about 85% of the cells express K14.
- 26. A population of cultured filaggrin- and involucrin-positive cells, wherein at least about 85% of the cells express filaggrin and involucrin.
Specification