Patent ID: 11913022
Assignee: CEDARS-SINAI MEDICAL CENTER
Field: Pharmaceuticals (Chemistry)
Classification: CPC C  A | IPC A  C

Claim 18:
19. 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;
differentiating the EBs into mammary cells comprising breast cells, luminal cells, and basal cells by culturing in a differentiation medium comprising hydrocortisone, insulin, FGF10, and HGF for 3D culture for about 28-32 days, wherein the EBs are cultured in the presence of pTHrP at a concentration of about 50 ng/ml-150 ng/ml from day 1 to day 5, the differentiation medium being a serum-free culture medium for the culture of mammary luminal and myoepithelial cells; and
differentiating the mammary cells to induce formation of the lactogenic mammary cells comprising culturing the mammary cells in the presence of insulin, prolactin and/or hydrocortisone.