Specification of functional cranial placode derivatives from human pluripotent stem cells

A method for inducing differentiation of cells, comprising

• a) contacting a plurality of cells with a first inhibitor of SMAD protein signaling (“first SMAD inhibitor”);
• b) contacting the cells with a second inhibitor of SMAD protein signaling (“second SMAD inhibitor”);
• and
• c) contacting the cells with a compound selected from the group consisting of BRL-54443, parthenolide, phenantroline, and combinations thereof,
• wherein the cells are contacted with the first SMAD inhibitor, the second SMAD inhibitor, and the compound in an amount effective to induce detectable expression of SIX1 and PAX6 in the plurality of cells.

A method of inducing differentiation of cells, comprising contacting a plurality of cells expressing SIX1 and PAX6 with

• (i) an activator of Wnt signaling (“Wnt activator”) in an amount effective to induce a detectable level of expression of SIX1 and PAX3 in the plurality of cells;
• (ii) with an activator sonic hedgehog (“SHH”) in an amount effective to induce a detectable level of expression of SIX1 and PITX1 in the plurality of cells;
• or
• (iii) with a bone morphogenetic protein (“BMP”) in an amount effective to induce a detectable level of expression of SIX1 and PITX3 in the plurality of cells.

A method of treating a subject comprising grafting a cell to a subject in need thereof, wherein

• a) the cell expresses a detectable level of SIX1 and PAX3, and the subject expresses at least one symptom of a pain syndrome, wherein grafting the cell reduces the at least one symptom of the pain syndrome;
• b) the cell expresses a detectable level of SIX1 and PITX1, and the subject expresses at least one symptom of a neuroendocrine disorder, wherein grafting the cell reduces the at least one symptom of the neuroendocrine disorder;
• or
• c) the cell expresses a detectable level of SIX1 and PITX3, and the subject expresses at least one symptom of an ocular disorder, wherein grafting the cell reduces the at least one symptom of the ocular disorder.