Abstract: Described are methods of enhancing development of renal organoids, methods of using the same, and kits.

Abstract: The Inventors established an efficient, chemically defined protocol for differentiating hPSCs into multi-potent nephron progenitor cells (NPCs) that can form nephron-like structures. By recapitulating metanephric kidney development in vitro, the Inventors generate SIX2+SALL1+WT1+PAX2+ NPCs with 90% efficiency within 9 days of differentiation. The NPCs possess the developmental potential of their in vivo counterparts and form PAX8+LHX1+ renal vesicles that self-pattern into nephron structures. In both 2D and 3D culture, NPCs form kidney organoids containing epithelial nephron-like structures expressing markers of podocytes, proximal tubules, loops of Henle, and distal tubules in an organized, continuous arrangement that resembles the nephron in vivo. The Inventors also show that this organoid culture system can be used to study mechanisms of human kidney development and toxicity.

Abstract: 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.