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US 12,351,822 B2Granted
Stem cell culture systems for columnar epithelial stem cells, and uses related thereto
AssigneeUniversity of Houston System
InventorsWa Xian, Frank McKeon, Matthew P. Vincent
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Abstract
The present invention relates to a culture media system that useful for the isolation and epigenetically stable propagation of normal stem cells in culture which are derived from columnar epithelial tissues and cancer stem cells from epithelial cancers. In certain embodiments, the culture system is a feeder-free system.
Claims12
A method for isolating a stem cell from columnar epithelial tissue comprising:
- (i) culturing dissociated epithelial cells from a columnar epithelial tissue sample to form stem cell colonies, wherein the dissociated cells and cell colonies are cultured in a medium comprising a ROCK (Rho Kinase) inhibitor, a Wnt agonist, a mitogenic growth factor, insulin (or an insulin mimetic) or IGF, a BRAF inhibitor, a VEGF inhibitor, nicotinamide, a Notch Agonist, a TGFβ signaling pathway inhibitor, and a Bone Morphogenetic Protein (BMP) antagonist;
- (ii) isolating single stem cells from the cell colonies, and
- (iii) culturing isolated single stem cells from step (ii) individually to form cultures purified stem cell clones;
- wherein each of the stem cell clones represents a clonal expansion of an epithelial stem cell present in the columnar epithelial tissue sample, thereby isolating epithelial stem cells.
1 more element
Dependencies are inferred from the claim text.