Methods for directed differentiation of pluripotent stem cells to immune cells

A method of producing immune cells comprising:

• (a) obtaining induced pluripotent stem cells (iPSCs), wherein the iPSCs are reprogrammed from a population of somatic cells;
• (b) incubating the iPSCs in a first defined media comprising a GSK inhibitor, wherein the media is free or essentially free of BMP4, IL-3, Flt3 ligand, and GM-CSF to prepare cells for HPC differentiation;
• (c) culturing the cells produced in step (b) in a second defined media comprising BMP4, FGF2, and VEGF to promote mesoderm induction in a plurality of the cells;
• (d) culturing the cells produced in step (c) in a third defined media comprising IL-3 and Flt3 ligand, such that a plurality of the cells proliferate and differentiate into HPCs that exhibit increased expression of one or more of CD7, DLL4, CD144, and CD235 as compared to cells produced in step (c);
• and
• (e) culturing the HPCs produced in step (d) under conditions to promote immune cell differentiation, thereby producing immune cells.