Selective In Vitro Propagation of Nephron Progenitors Derived from Embryos and Pluripotent Stem Cells

•Purified SIX2+ murine nephron progenitors are selectively expanded in vitro•Expanded SIX2+ cells reconstitute 3D glomeruli and renal tubules•Mouse ESC-derived nephron progenitors are expanded with nephron-forming potential•Human iPSC-derived nephron progenitors are propagated and form nephrons

SummaryNephron progenitors in the embryonic kidney propagate while generating differentiated nephrons. However, in mice, the progenitors terminally differentiate shortly after birth. Here, we report a method for selectively expanding nephron progenitors in vitro in an undifferentiated state. Combinatorial and concentration-dependent stimulation with LIF, FGF2/9, BMP7, and a WNT agonist is critical for expansion. The purified progenitors proliferated beyond the physiological limits observed in vivo, both for cell numbers and lifespan. Neonatal progenitors were maintained for a week, while progenitors from embryonic day 11.5 expanded 1,800-fold for nearly 20 days and still reconstituted 3D nephrons containing glomeruli and renal tubules. Furthermore, progenitors generated from mouse embryonic stem cells and human induced pluripotent cells could be expanded with retained nephron-forming potential. Thus, we have established in vitro conditions for promoting the propagation of nephron progenitors, which will be essential for dissecting the mechanisms of kidney organogenesis and for regenerative medicine.

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