Generation of kidney organoids derived from human expanded potential stem cells.

The establishment of human expanded potential stem cell (hEPSC) presents a unique cellular platform for translational research in kidney organoids. We generated SIX2 reporter and doxycycline (DOX)-inducible YAP overexpression in hEPSC lines using CRISPR-Cas9. Chemical compounds and DOX were added to the culture medium to induce Hippo-YAP signaling, respectively. The hEPSC line containing the SIX2-mCherry reporter gene accurately reflected SIX2 expression in vitro, enabling the real-time tracking of kidney organoid development. A comparative analysis revealed that inhibiting the Hippo-YAP signaling pathway before nephron progenitor cell (NPC) generation effectively increased the number of NPCs, resulting in a more nephron-like structure. However, prolonged inhibition hindered the further maturation of the kidney organoids, leading to differentiation stagnation. Therefore, activating YAP before NPC generation facilitates their maturation, offering effective induction strategies improving kidney organoid differentiation efficiency.