	US 12,203,099 B2
	Methods of expanding cholangiocytes
Ludovic Vallier, Cambridge (GB); Nicholas Hannan, Cambridge (GB); Kourosh Saeb-Parsy, Cambridge (GB); and Fotios Sampaziotis, Cambridge (GB)
Assigned to CAMBRIDGE ENTERPRISE LIMITED, Cambridge (GB)
Appl. No. 16/623,701
Filed by Cambridge Enterprise Limited, Cambridge (GB)
PCT Filed Jun. 19, 2018, PCT No. PCT/EP2018/066295 § 371(c)(1), (2) Date Dec. 17, 2019, PCT Pub. No. WO2018/234323, PCT Pub. Date Dec. 27, 2018.
Claims priority of application No. 1709704 (GB), filed on Jun. 19, 2017.
Prior Publication US 2021/0139856 A1, May 13, 2021
Int. Cl. C12N 5/071 (2010.01); A61K 35/413 (2015.01); A61P 1/16 (2006.01)

CPC C12N 5/0679 (2013.01) [A61K 35/413 (2013.01); A61P 1/16 (2018.01); C12N 2501/11 (2013.01); C12N 2501/415 (2013.01); C12N 2513/00 (2013.01); G01N 2800/52 (2013.01)]

9 Claims

1. 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.