A developmentally-inspired hypoxia condition promotes kidney organoids differentiation from human pluripotent stem cells

Kidney organoids derived from human pluripotent stem cells (hPSCs) lack a contiguous network of collecting ducts, which limits their utility in modeling kidney development and disease. Here, we report the generation of kidney organoids containing ureteric bud (UB)-derived collecting ducts connected to metanephric mesenchyme (MM)-derived nephrons using a developmentally-inspired hypoxic differentiation condition. Hypoxia promotes a reiterative process of branching morphogenesis and nephron induction through reciprocal interactions between co-induced MM and UB, which lead to a higher-order kidney organogenesis in vitro. The resulting kidney organoids demonstrate greater maturity, as indicated by higher levels of functional markers and a more realistic micro-anatomy. Additionally, these hypoxic-enhanced kidney organoids show a great potential as in vitro models for renal cystic diseases, as they efficiently generate cystic formations and display high sensitivity to drugs. This hypoxia approach may open new avenues for an enhanced understanding of kidney development and diseases. Overall design: Single-cell RNA sequencing was conducted to profile the cellular composition of kidney organoids differentiated under normoxic and hypoxic conditions.