Metabolic Substrate Availability Regulates Nephron Lineage Expansion in Human Kidney Organoids

ABSTRACT: Human induced pluripotent stem cell-derived kidney organoids are widely used to model nephrogenesis, but variability in basal media composition remains an underappreciated source of developmental heterogeneity. Because nephrogenesis involves rapid proliferation and differentiation, we hypothesized that metabolic substrate availability directly regulates organoid development. We compared growth and cellular composition across commonly used media (APEL2, advanced RPMI, and DMEM). Advanced RPMI supported robust growth and tubular expansion, whereas DMEM with low basal glucose (5.0 mM) impaired nephron lineage development. Glucose supplementation (to 11.1 mM) significantly rescued these defects, indicating a causal role for substrate limitation. Metabolomics analysis further revealed rapid glucose depletion under low glucose conditions and reduced metabolic activity, while higher glucose media supported oxidative metabolism. Functionally, glucose availability directly influenced cellular proliferation and survival, with low glucose associated with increased TUNEL positivity and cleaved caspase-3. Together, these findings identify glucose as a critical regulator of kidney organoid development and position metabolic substrate composition as a tunable determinant of organoid fidelity and reproducibility.

INSTRUMENT(S): Nuclear Magnetic Resonance (NMR) -

PROVIDER: MTBLS16048 | MetaboLights | 2026-10-18

REPOSITORIES: MetaboLights