Described herein are methods related to generating intermediate
mesoderm (IM) cells, including using
sequential treatment of small molecules and growth factors, and composition produced by the described methods. Using small molecules such as CHIR99021 in combination with FGF2 and RA, efficient differentiation of human pluripotent stem cells (hPSCs) into intermediate
mesoderm, such as PAX2+LHX1+ cells, is achieved. The method is extensible different hPSC
cell lines and does not require
flow sorting. Importantly, resulting PAX2+LHX1+ cells, are capable of WT1 expression and addition of FGF9 and activin, PAX2+LHX1+ cells specifically differentiates cells into SIX2, SALL1, and WT1 expressing cells representative of cap
mesenchyme nephron progenitor cells. The described methods and compositions facilitate and improve the
directed differentiation of hPSCs into cells of the
kidney lineage for the purposes of bioengineering
kidney tissue and iPS
cell disease modeling.