Methods for directed differentiation of pluripotent stem cells to HLA homozygous immune cells

A method of producing HLA homozygous lymphoid cells comprising:

• (a) obtaining induced pluripotent stem cells (iPSCs), wherein the iPSCs have been reprogrammed from a population of HLA homozygous blood cells;
• (b) incubating the iPSCs in a first defined media comprising a GSK inhibitor, wherein the media is essentially free of BMP4, IL-3, Flt3 ligand, and GM-CSF to produce cells in the first stage of HPC differentiation;
• (c) culturing the cells in the first stage HPC differentiation in a second defined media comprising BMP4, FGF2, and VEGF to produce cells in the second stage of HPC differentiation;
• (d) culturing the cells in the second stage of HPC differentiation in a third defined media comprising IL-3 and Flt3 ligand, such that a plurality of the cells proliferate and differentiate into HPCs;
• (e) measuring the expression of CD7, DLL4, CD144, and CD235 in the HPCs during HPC differentiation to identify cells that exhibit increased expression of CD7, DLL4, CD144, and CD235 as compared to cells in the second stage of HPC differentiation;
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• (f) enriching the HPCs that exhibit increased expression of CD7, DLL4, CD144,and CD235 for CD34+/CD45+/CD7+/CD144+HPCs;
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• (g) culturing the CD34+/CD45+/CD7+/CD144+ HPCs under conditions to promote lymphoid cell differentiation, thereby producing HLA homozygous lymphoid cells.