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US 12,203,099 B2Granted
Methods of expanding cholangiocytes
AssigneeCambridge Enterprise Limited
InventorsLudovic Vallier, Nicholas Hannan, Kourosh Saeb-Parsy, Fotios Sampaziotis
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Abstract
This invention relates to the expansion of primary cholangiocytes in the form of cholangiocyte organoids (COs) using culture conditions in which canonical Wnt signalling is inhibited and non-canonical Wnt/PCP signalling is potentiated. Methods of expanding primary cholangiocytes, expanded populations of cholangiocytes and medical applications of expanded cholangiocytes are provided.
Claims9
A method for expanding human primary cholangiocytes in vitro comprising:
- (i) providing a population of isolated human primary cholangiocytes:
- (ii) forming cholangiocyte organoids from the population of isolated human primary cholangiocytes and;
- (iii) culturing the cholangiocyte organoids in an expansion medium in conditions sufficient for expanding human primary cholangiocytes, to produce an expanded population of human primary cholangiocytes,
- wherein the expansion medium is a nutrient medium supplemented with growth factors comprising epidermal growth factor (EGF), a canonical Wnt signaling inhibitor and R-spondin;
- and
- the canonical Wnt signaling inhibitor increases the phosphorylation of B-catenin in the cholangiocyte organoids.
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Dependencies are inferred from the claim text.