	US 11,913,022 B2
	In vitro induction of mammary-like differentiation from human pluripotent stem cells
Ying Qu, Los Angeles, CA (US); Xiaojiang Cui, Arcadia, CA (US); Dhruv Sareen, Porter Ranch, CA (US); and Armando E. Giuliano, Los Angeles, CA (US)
Assigned to Cedars-Sinai Medical Center, Los Angeles, CA (US)
Appl. No. 16/480,778
Filed by Cedars-Sinai Medical Center, Los Angeles, CA (US)
PCT Filed Jan. 25, 2018, PCT No. PCT/US2018/015318 § 371(c)(1), (2) Date Jul. 25, 2019, PCT Pub. No. WO2018/140647, PCT Pub. Date Aug. 2, 2018.
Claims priority of provisional application 62/450,484, filed on Jan. 25, 2017.
Prior Publication US 2020/0002671 A1, Jan. 2, 2020
Int. Cl. A61K 35/55 (2015.01); C12N 5/071 (2010.01); C12N 5/074 (2010.01)

CPC C12N 5/0631 (2013.01) [A61K 35/55 (2013.01); C12N 5/0696 (2013.01); C12N 2506/45 (2013.01); C12N 2533/54 (2013.01); C12N 2533/90 (2013.01)]

19 Claims

1. A method of generating mammary cells, comprising:

culturing human induced pluripotent stem cells (hiPSCs) in a serum-free culture medium enriching non-neural ectoderm cells, the serum-free culture medium comprising a basal medium, proliferation supplements, heparin, and hydrocortisone for the culture of non-neural ectoderm cell-containing mammospheres, for about 8-12 days to generate embryoid bodies (EBs) expressing non-neural ectoderm stem cell markers AP-2γ, CK8 and CK18, wherein the EBs do not express neural ectoderm stem cell markers OTX and SOX11; and

differentiating the EBs into mammary cells comprising breast cells, luminal cells, and basal cells by culturing in a differentiation medium supplemented with pTHrP from day 1 to day 5, followed by culturing in the differentiation medium supplemented with hydrocortisone, insulin, FGF10, and HGF for 3D culture for about 23-27 days, the differentiation medium being a serum-free culture medium for the culture of mammary luminal and myoepithelial cells.