Patent ID: 9624468
Date: 2017-04-18
CPC Classifications: C12N,G01N

Claim:
1. A method for producing preplacodal ectoderm cells, the method comprising the steps of: (i) culturing pluripotent stem cells selected from the group consisting of human pluripotent stem cells and mouse pluripotent stem cells for between 1 to 2 days under conditions that result in formation of embryoid bodies from the cultured pluripotent stem cells; (ii) adding one of (1) a gelatinous protein mixture secreted by Engelbreth-Holm-Swarm (EHS) mouse sarcoma cells or (2) an effective amount of laminin and entactin, wherein the effective amount facilitates the formation of a basement membrane, to the cultured embryoid bodies formed in step (i); and (iii) culturing the embryoid bodies of step (ii) for about 2 days to about 3 days in the presence of BMP2, BMP4, or BMP7 and a TGF-β inhibitor to form non-neural ectoderm cells; and (iv) culturing the non-neural ectoderm cells formed in (iii) for about 3 days to about 5 days in the absence of the BMP2, BMP4, or BMP7 and the TGF-β inhibitor, and in the presence of an exogenous FGF and a BMP inhibitor, in floating culture for about 4 days to about 6 days after initiation of step (i), to generate a cell population comprising human or mouse preplacodal ectoderm cells, wherein said human preplacodal ectoderm cells express one or more of SIX1/4, EYA1/2, IRX1/2/3, AP2, DLX3/5/6, GATA2/3, FOXI1/3 and E-cadherin or wherein said mouse preplacodal ectoderm cells express at least one of Dlx3, Dlx5, Pax2, Pax8 and Eya1.